Thlaspi Caerulescens Research Articles

Phytoextraction of cadmium and zinc from arable soils amended with sewage sludge using Thlaspi caerulescens: Development of a predictive model

The objectives of this study were to assess the potential for using Thlaspi caerulescens as a phytoextraction plant and develop a user-advice model, which can predict the frequency of phytoextraction operation required under prescribed conditions. Pot and field trials were conducted using soil collected from a dedicated sewage sludge disposal facility. Soil amendments (sulphuric acid, potassium chloride and EDTA) intended to increase Cd solubility were also tested. Predictive models of Cd and Zn uptake were developed which were able to reproduce the observed pH-dependence of Cd uptake with an apparent maximum around pH 6. Chemical treatments did not significantly increase the uptake of Cd. The periodic use of phytoextraction with T. caerulescens to maintain soils below statutory metal concentration limits, when modern sewage sludges are repeatedly applied, seems very attractive given the non-intrusive and cost-effective nature of the process. The major limitations lie with the large-scale husbandry of T. caerulescens.

A method for cellular localization of gene expression via quantitative in situ hybridization in plants

A quantitative in situ hybridization technique (quantitative whole-mount in situ hybridization, QISH) for plants is described. It employs direct hybridization of fluorescently labelled gene-specific oligonucleotides in large tissue pieces combined with optical sectioning. It dramatically increases the throughput compared with conventional antibody- and microtome-based in situ mRNA hybridization methods, while simultaneously eliminating artefact-prone preparation steps that prevent reliable quantification in conventional methods. The key feature of this technique is the quantification of gene expression using housekeeping genes (cytosolic GAPDH and 18S RNA) as internal standards. This feature enables a correction of varying cytoplasm/vacuole ratios in different cell types, as well as tissue optical effects and non-specific signals. The quantitative nature of the technique allows for analysis of gene expression in response to different environmental conditions, as well as tissue- and age-dependent differences in gene expression patterns. In addition to testing tissue permeabilization, structural preservation, specificity, linearity and tissue optical effects, we verified the reliability of the technique with three Arabidopsis thaliana genes of known function and distribution. These were the rbcL gene for ribulose 1,5-bisphosphate carboxylase, the developmentally related gene SCARECROW (AtSCR) and PHOT-1, a photoreceptor kinase. As expected, rbcL mRNA was found in all photosynthetic cells, while SCR mRNA was detected mainly in bundle sheath cells and PHOT-1 was found predominantly in epidermal and cortical cells of the apical hook of light-grown seedlings. As an application example, QISH was used to measure transcript abundance for a zinc transporter from the ZIP family of transporters in the Zn/Cd hyperaccumulator model plant, Thlaspi caerulescens, and the related non-accumulator Thlaspi arvense. This showed that QISH can be used to compare differences in mRNA levels between cell types, plant growth conditions and plant species. Messenger RNA for the zinc transporter gene ZNT1 was abundant in photosynthetic cells, but not in the epidermal storage cells where metal hyperaccumulation in T. caerulescens occurs. This indicates that ZNT1 does not directly participate in metal hyperaccumulation within the leaf. Growing T. caerulescens with high zinc levels strongly reduced ZNT1 transcript abundance in the spongy mesophyll cells, but less in the other cell types. In T. arvense, ZNT1 mRNA levels were generally much lower, and were furthermore drastically reduced by growth at increased zinc levels, confirming earlier reports regarding ZNT1 regulation in these two Thlaspi species.

Soil Microbial Diversity as Affected by the Rhizosphere of the Hyperaccumulator Thlaspi Caerulescens Under Natural Conditions

It is hypothesized that metal hyperaccumulator plants have specific rhizosphere conditions, potentially modifying the bioavailability of soil metals. This article aims to further the knowledge about the rhizosphere of the hyperaccumulator Thlaspi caerulescens, focusing on its microflora isolated from metalliferous soils collected in situ where the plants grow naturally. We characterized the cultivable microbial communities isolated from the rhizosphere of one population of this Ni hyperaccumulator species grown on a serpentine soil. The rhizosphere soil harbored a wide variety of microorganisms, predominantly bacteria, confirming the stimulatory effect of the T. caerulescens rhizosphere on microbial growth and proliferation. We tested the hypothesis that the rhizosphere of T. caerulescens influences 1) the metabolic diversity of the bacterial community and 2) the bacterial resistance to metals. The principal component analysis of the Biolog® plate's data confirmed a structural effect of the rhizosphere of T. caerulescens on bacterial communities. The percentage of Ni-resistant bacteria was higher in the rhizosphere than in the bulk soil, suggesting a direct effect of the rhizosphere on Ni tolerance, reflecting a greater bacterial tolerance to Ni in the rhizosphere.

Do metal-rich plants deter herbivores? A field test of the defence hypothesis

Some plant species growing on metalliferous soils are able to accumulate heavy metals in their shoots up to very high concentrations, but the selective advantage of this behaviour is still unknown. The most popular hypothesis, that metals protect plants against herbivores, has been tested several times in laboratory conditions, with contradictory results. We carried out the first large-scale test of the defence hypothesis in eight natural populations of the model Zn hyperaccumulator Thlaspi caerulescens J. and C. Presl (Brassicaceae). In two climatic regions (temperate, Belgium-Luxembourg, and Mediterranean, southern France), we worked in metalliferous and in normal, uncontaminated environments, with plants spanning a wide range of Zn concentrations. We also examined the importance of glucosinolates (main secondary metabolites of Brassicaceae) as antiherbivore defences. When exposed to natural herbivore populations, T. caerulescens suffered lower herbivory pressures in metal-enriched soils than in normal soils, both in Belgium-Luxembourg and in southern France. The trapping of gastropods shows an overall lower population density in metalliferous compared to normal environments, which suggests that herbivory pressure from gastropods is lower on metalliferous soils. In addition, foliar concentration of glucosinolates was constitutively lower in all populations from metal-enriched soils, suggesting that these have evolved towards lower investment in organic defences in response to lower herbivory pressure. The Zn concentration of plants had a protective role only for Belgian metallicolous plants when transplanted in normal soils of Luxembourg. These results do not support the hypothesis that Zn plays a key role in the protection of T. caerulescens against enemies. In contrast, glucosinolates appear to be directly involved in the defence of this hyperaccumulator against herbivores.

Predicting the Phytoextraction Duration to Remediate Heavy Metal Contaminated Soils

The applicability of phytoextraction to remediate soils contaminated with heavy metals (HMs) depends on, amongst others, the duration before remediation is completed. The impact of changes in the HM content in soil occurring during remediation on plant uptake has to be considered in order to obtain a reliable estimate of the phytoextraction duration. To simulate the decrease in the HM content in soil and to assess the resulting decrease in the uptake of HMs by plants, contaminated soil was mixed with uncontaminated, but otherwise similar soil. Uptake of Cd, Pb, and Zn by the indicator plant Lupinus hartwegii and the Zn hyperaccumulator Thlaspi caerulescens (La Calamine ecotype) was a log-linear function of the in-situ measured HM soil solution concentrations. Over a wide range in dissolved Cd and Zn concentrations, uptake of these HMs by T. caerulescens was (much) greater than by L. hartwegii. Experimentally derived regression models describing the relationships between soil, soil solution, and plant were implemented in a HM mass balance model used to obtain estimates of the phytoextraction duration. For our target soils, estimates of the Cd phytoextraction duration using L. hartwegii or T. caerulescens increased significantly by more than 100 or 50 years when experimental soil—soil solution—plant relationships were used instead of the assumption of constant plant uptake of Cd. The two approaches gave similar results for phytoextraction of Zn by T. caerulescens.

Testing of Outstanding Individuals of Thlaspi Caerulescens for Cadmium Phytoextraction

Thlaspi caerulescens is known to hyperaccumulate high quantities of Cd with Cd concentrations up to 3000 mg kg−1 in some populations from south of France. However, within these populations, the Cd concentrations can vary widely from plant to plant in a way that appears to be not entirely due to variations in soil Cd. The aim of this work was to investigate the variability in the Cd uptake ability of individual plants within a population and among seedlings grown from seeds from a single plant. Ten populations of T. caerulescens plants were selected from four locations (V: Viviez; SF: Saint Félix-de-Pallières; LB: Le Bleymard; CMA: Col du Mas de l’Air) depending of the extent and soil homogeneity of the site. One population from CMA consisted of the progeny of a single maternal plant. Hundred plants of each population were grown for three months in the same homogeneous and lightly Cd-polluted soil (about 20 mg total Cd kg−1 dry soil). Cadmium uptake behavior of the plants was monitored by labeling the soil with 109Cd. To allow partial plant destruction, radioanalysis was performed on the largest leaf of each plant as an indicator of the total Cd concentration in plant shoots. Results showed significant differences in biomass production and Cd uptake by T. caerulescens between sites and between populations within sites. We observed a wide intra-population variation in biomass, Cd concentration and total Cd uptake. For these properties, 1 to 5 percents of the plants in each population varied by more than a factor of two from the mean values. The mean Cd uptake by the single-plant population from CMA was more than 40% higher than for the population at large. T. caerulescens would respond to traditional selection methods, which would significantly improve the phytoextraction of Cd.

TcYSL3, a member of the YSL gene family from the hyper‐accumulator Thlaspi caerulescens, encodes a nicotianamine‐Ni/Fe transporter

The two main features of plant hyper-accumulator species are the massive translocation of heavy metal ions to the aerial parts and their tolerance to such high metal concentrations. Recently, several lines of evidence have indicated a role for nicotianamine (NA) in metal homeostasis, through the chelation and transport of NA-metal complexes. The function of transport of NA-metal chelates, required for the loading and unloading of vessels, has been assigned to the Yellow Stripe 1 (YSL)-Like family of proteins. We have characterized three YSL genes in Thlaspi caerulescens in the context of hyper-accumulation. The three YSL genes are expressed at high rates compared with their Arabidopsis thaliana homologs but with distinct patterns. While TcYSL7 was highly expressed in the flowers, TcYSL5 was more highly expressed in the shoots, and the expression of TcYSL3 was equivalent in all the organs tested. In situ hybridizations have shown that TcYSL7 and TcYSL5 are expressed around the vasculature of the shoots and in the central cylinder in the roots. The exposure to heavy metals (Zn, Cd, Ni) does not affect the high and constitutive expression of the TcYSL genes. Finally, we have demonstrated by mutant yeast complementation and uptake measurements that TcYSL3 is an Fe/Ni-NA influx transporter. This work provides therefore molecular, histological and biochemical evidence supporting a role for YSL transporters in the overall scheme of NA and NA-metal, particularly NA-Ni, circulation in a metal hyper-accumulator plant.

Root-to-shoot long-distance circulation of nicotianamine and nicotianamine-nickel chelates in the metal hyperaccumulator Thlaspi caerulescens

Plant metal hyperaccumulator species are widely used as models to unravel the heavy metal tolerance and hyperaccumulation mechanisms. Thlaspi caerulescens is capable of tolerating and hyperaccumulating Zn, Cd, and Ni. A search for factors involved in the cellular tolerance to Ni, based on yeast screens, led to isolation of a cDNA encoding a functional nicotianamine (NA) synthase (NAS). The T. caerulescens genome appears to contain a single copy of the NAS gene named TcNAS whose expression is restricted to the leaves. The analysis of dose-response and time-course Ni treatments have revealed that the exposure to Ni triggers the accumulation of NA in the roots. Because neither TcNAS expression nor NAS activity were detected in the roots, the NA accumulation in roots is most probably the result of its translocation from the leaves. Once in the roots, NA, together with Ni, is subsequently found in the xylem, for redirection to the aerial parts. Using liquid chromatography coupled to inductively coupled plasma or electrospray ionization mass spectrometry, it has been shown that part of the Ni is translocated as a stable Ni-NA complex in the xylem sap. This circulation of NA, Ni, and NA-Ni chelates is absent in the non-tolerant non-hyperaccumulator related species T. arvense. Taken together, the results provide direct physiological and chemical evidence for NA and NA-heavy metal complex translocation in a hyperaccumulator species.

Life history variation in the heavy metal tolerant plant Thlaspi caerulescens growing in a network of contaminated and noncontaminated sites in southern France: role of gene flow, selection and phenotypic plasticity

* Here we explore life history differences in a set of neighbouring metallicolous and nonmetallicolous populations of the heavy metal tolerant plant Thlaspi caerulescens. * We contrasted data from field observations and from a common garden experiment, in which soil zinc (Zn) concentration and light availability were manipulated, and data on microsatellite molecular variation. * The two ecotypes showed few differences in life history in the field, but large differences in their response to Zn concentration in the common garden. Soil toxicity affected most characters in nonmetallicolous plants, while it had no effect on metallicolous plants. The two ecotypes responded similarly to light. Genetic differentiation for quantitative characters between ecotypes contrasted with the absence of differentiation for microsatellites. Conversely, populations of the same ecotype showed similar responses to Zn, despite their high differentiation for molecular markers. * We conclude that divergent selection related to soil toxicity has had a predominant role in shaping life history differences between ecotypes, gene flow weakly opposing local adaptation despite geographical proximity.

Isolation of Zn-responsive genes from two accessions of the hyperaccumulator plant Thlaspi caerulescens

Several populations with different metal tolerance, uptake and root-to-shoot transport are known for the metal hyperaccumulator plant Thlaspi caerulescens. In this study, genes differentially expressed under various Zn exposures were identified from the shoots of two T. caerulescens accessions (calaminous and non-calaminous) using fluorescent differential display RT-PCR. cDNA fragments from 16 Zn-responsive genes, including those encoding metallothionein (MT) type 2 and type 3, MRP-like transporter, pectin methylesterase (PME) and Ole e 1-like gene as well as several unknown genes, were eventually isolated. The full-length MT2 and MT3 sequences differ from those previously isolated from other Thlaspi accessions, possibly representing new alleles or isoforms. Besides the differential expression in Zn exposures, the gene expression was dependent on the accession. Thlaspi homologues of ClpP protease and MRP transporter were induced at high Zn concentrations. MT2 and PME were expressed at higher levels in the calaminous accession. The MTs and MRP transporter expressed in transgenic yeasts were capable of conferring Cu and Cd tolerance, whereas the Ole e 1-like gene enhanced toxicity to these metals. The MTs increased yeast intracellular Cd content. As no significant differences were found between Arabidopsis and Thlaspi MTs, they apparently do not differ in their capacity to bind metals. However, the higher levels of MT2 in the calaminous accession may contribute to the Zn-adapted phenotype.

Reaction norms of life history traits in response to zinc in Thlaspi caerulescens from metalliferous and nonmetalliferous sites

* We examined phenotypic plasticity of fitness components in response to zinc (Zn) in the Zn hyperaccumulator, Thlaspi caerulescens. * Two populations from Zn-enriched soils (M) and two populations from normal soils (NM) were grown in pots at three Zn concentrations (0, 1000 and 8000 mg kg(-1) Zn), for an entire life cycle. Growth, Zn accumulation and fitness components were assessed. * Based on vegetative growth, M and NM populations had similar Zn tolerance at 1000 mg kg(-1) Zn. However, reproductive output was markedly decreased in NM at 1000 and 8000 mg kg(-1) Zn. In M populations, Zn did not affect fitness. However, low Zn status enhanced reproductive output in year 1 compared with year 2 and decreased survival after the first flowering season. * M populations are able to achieve equal fitness across a broad range of Zn concentrations in soil by different combinations of fecundity and longevity. No cost of higher tolerance was demonstrated in M populations. Reproductive traits appeared to be a more sensitive indicator of tolerance than vegetative growth.

QTL analysis of cadmium and zinc accumulation in the heavy metal hyperaccumulator Thlaspi caerulescens

Thlaspi caerulescens (Tc; 2n = 14) is a natural Zn, Cd and Ni hyperaccumulator species belonging to the Brassicaceae family. It shares 88% DNA identity in the coding regions with Arabidopsis thaliana (At) (Rigola et al. 2006). Although the physiology of heavy metal (hyper)accumulation has been intensively studied, the molecular genetics are still largely unexplored. We address this topic by constructing a genetic map based on AFLP markers and expressed sequence tags (ESTs). To establish a genetic map, an F(2) population of 129 individuals was generated from a cross between a plant from a Pb/Cd/Zn-contaminated site near La Calamine, Belgium, and a plant from a comparable site near Ganges (GA), France. These two accessions show different degrees of Zn and, particularly, Cd accumulation. We analyzed 181 AFLP markers (of which 4 co-dominant) and 13 co-dominant EST sequences-based markers and mapped them to seven linkage groups (LGs), presumably corresponding to the seven chromosomes of T. caerulescens. The total length of the genetic map is 496 cM with an average density of one marker every 2.5 cM. This map was used for Quantitative Trait Locus (QTL) mapping in the F(2). For Zn as well as Cd concentration in root we mapped two QTLs. Three QTLs and one QTL were mapped for Zn and Cd concentration in shoot, respectively. These QTLs explain 23.8-60.4% of the total variance of the traits measured. We found only one common locus (LG6) for Zn and Cd (concentration in root) and one common locus for shoot and root concentrations of Zn (LG1) and of Cd (LG3). For all QTLs, the GA allele increased the trait value except for two QTLs for Zn accumulation in shoot (LG1 and LG4) and one for Zn concentration in root (LG1).

Synthesis of low molecular weight thiols in response to Cd exposure in Thlaspi caerulescens

In this study, we investigated the accumulation of phytochelatins (PCs) and other low molecular weight (LMW) thiols in response to Cd exposure in two contrasting ecotypes differing in Cd accumulation. Using a root elongation test, we found that the highly accumulating ecotype Ganges was more tolerant to Cd than the low Cd-accumulation ecotype Prayon. L-buthionine-(S,R)-sulphoximine (BSO), a potent inhibitor of the gamma-glutamylcysteine synthetase gamma-ECS) (an enzyme involved in the PC biosynthetic pathway), increased the Cd sensitivity of Prayon, but had no effect on Ganges. Although PC accumulation increased in response to Cd exposure, no significant differences were observed between the two ecotypes. Cd exposure induced a dose-dependent accumulation of both Cys and a still unidentified LMW thiol in roots of both ecotypes. Root accumulation of Cys and this thiol was higher in Ganges than in Prayon; the ecotypic differences were more pronounced when the plants were treated with BSO. These findings suggest that PCs do not contribute to the Cd hypertolerance displayed by the Ganges ecotype of Thlaspi caerulescens, whereas Cys and other LMW thiols might be involved.

The heavy metal hyperaccumulator Thlaspi caerulescens expresses many species‐specific genes, as identified by comparative expressed sequence tag analysis

Thlaspi caerulescens is a natural zinc (Zn), cadmium (Cd) and nickel (Ni) hyperaccumulator and an emerging plant model species to study heavy metal hyperaccumulation and tolerance. This paper describes the analysis of the first expressed sequence tag (EST) collection from T. caerulescens. This collection is a new resource to unravel the molecular basis of plant metal homeostasis, tolerance and hyperaccumulation. In total, 4289 ESTs were generated originating from Zn-exposed root and shoot tissues, leading to 3709 T. caerulescens assembled partial cDNA sequences (unigenes). In comparison to Arabidopsis or other publicly available plant sequences, a fraction of c. 8% of the T. caerulescens unigenes (TcUGs) had no significant similarity with any known DNA sequence and, so far, these sequences are T. caerulescens specific. Three per cent of the TcUGs correspond to Arabidopsis thaliana orthologues that, as yet, have not been found to be expressed. The T. caerulescens transcriptome generally relates very well to the A. thaliana transcriptome, although, compared with other closely related species, a relatively large number of T. caerulescens-specific transcripts were found. T. caerulescens also expresses a relatively large number of genes which are expressed at a very low level in A. thaliana.

Analysis of Micro-Nutrient Behaviour in the Rhizosphere using a DGT Parameterised Dynamic Plant Uptake Model

Mathematical models of micro-nutrient uptake by plants have been based on models designed for uptake of major nutrients. This poses a number of problems, as the behaviour of micro-nutrients in the rhizosphere is different, particularly with respect to interactions between solid and solution compartments. There is no standard procedure for the measurement of soil parameters that affect the supply of micro-nutrient from the soil. The Dynamic Plant Uptake Model (DPUM) presented here can use soil parameters for micro-nutrients that are measured under similar conditions to those that apply during plant uptake, as, for example, by the technique of DGT (Diffusive Gradients in Thin-films). The construction and practical applications of the model are discussed in the context of mathematical simulations of realistic situations. The simulations have been used to investigate conditions under which uptake of zinc by a zinc hyperaccumulator (Thlaspi caerulescens), and a non-accumulator plant (Thlaspi arvense), is limited by either processes occurring in the plant or in the soil. The size of the solid phase reservoir of zinc, and the rate at which the solid phase reservoir responds to zinc depletion in the solution phase, are important in determining the soil solution concentration at which the plant uptake kinetics begin to limit the uptake of the metal. The relative importance of advective transport of zinc from the surrounding soil to the root–soil interface depends on both the size of the solid phase reservoir and the speed at which the soil responds to zinc depletion in the pore water. These results have been used to assess conditions where DGT is likely to behave as a reliable surrogate for the transport and metal release processes that affect plant uptake.

A comparison of phytoremediation capability of selected plant species for given trace elements

In our experiment, As, Cd, Pb, and Zn remediation possibilities on medium contaminated soil were investigated. Seven plant species with a different trace element accumulation capacity and remediation potential were compared. We found good accumulation capabilities and remediation effectiveness of Salix dasyclados similar to studied hyperaccumulators (Arabidopsis halleri and Thlaspi caerulescens). We have noticed better remediation capability in willow compared to poplar for most of the elements considered in this experiment. On the contrary, poplar species were able to remove a larger portion of Pb as opposed to other species. Nevertheless, the removed volume was very small. The elements found in plant biomass depend substantially on the availability of these elements in the soil. Different element concentrations were determined in natural soil solution and by inorganic salt solution extraction (0.01molL−1 CaCl2). Extracted content almost exceeded the element concentration in the soil solution. Element concentrations in soil solution were not significantly affected by sampling time.

Soil pH Effects on Uptake of Cd and Zn by Thlaspi caerulescens

For phytoextraction to be successful and viable in environmental remediation, strategies that can optimize plant uptake must be identified. Thlaspi caerulescens is an important hyperaccumulator of Cd and Zn, whether adjusting soil pH is an efficient way to enhance metal uptake by T. caerulescens must by clarified. This study used two soils differing in levels of Cd and Zn, which were adjusted to six different pH levels. Thlaspi caerulescens tissue metal concentrations and 0.1 M Sr(NO3)2 extractable soil metal concentrations were measured. The soluble metal form of both Cd and Zn was greatly increased with decreasing pH. Lowering pH significantly influenced plant metal uptake. For the high metal soil, highest plant biomass was at the lowest soil pH (4.74). The highest shoot metal concentration was at the second lowest pH (5.27). For low metal soil, due to low pH induced Al and Mn toxicity, both plant growth and metal uptake was greatest at intermediate pH levels. The extraordinary Cd phytoextraction ability of T. caerulescens was further demonstrated in this experiment. In the optimum pH treatments, Thlaspi caerulescens extracted 40% and 36% of total Cd in the low and high metal soils, respectively, with just one planting. Overall, decreasing pH is an effective strategy to enhance phytoextraction. But different soils had various responses to acidification treatment and a different optimum pH may exist. This pH should be identified to avoid unnecessarily extreme acidification of soils.

A comparison of the Thlaspi caerulescens and Thlaspi arvense shoot transcriptomes

Whole-genome transcriptome profiling is revealing how biological systems are regulated at the transcriptional level. This study reports the development of a robust method to profile and compare the transcriptomes of two nonmodel plant species, Thlaspi caerulescens, a zinc (Zn) hyperaccumulator, and Thlaspi arvense, a nonhyperaccumulator, using Affymetrix Arabidopsis thaliana ATH1-121501 GeneChip arrays (Affymetrix, Santa Clara, CA, USA). Transcript abundance was quantified in the shoots of agar- and compost-grown plants of both species. Analyses were optimized using a genomic DNA (gDNA)-based probe-selection strategy based on the hybridization efficiency of Thlaspi gDNA with corresponding A. thaliana probes. In silico alignments of GeneChip probes with Thlaspi gene sequences, and quantitative real-time PCR, confirmed the validity of this approach. Approximately 5000 genes were differentially expressed in the shoots of T. caerulescens compared with T. arvense, including genes involved in Zn transport and compartmentalization. Future functional analyses of genes identified as differentially expressed in the shoots of these closely related species will improve our understanding of the molecular mechanisms of Zn hyperaccumulation.

Fixation of metals in soil constituents and potential remobilization by hyperaccumulating and non-hyperaccumulating plants: Results from an isotopic dilution study

In this study isotopic dilution methods were used to investigate the hypothesis that access to metals associated with specific chemical components in the soil that are not available to non-accumulator species could be involved in hyperaccumulation. The hyperaccumulator Thlaspi caerulescens and a non-accumulator species, Brassica napus, were grown in Cd and Zn enriched soil components calcite, goethite, charcoal and cryptomelane. The metal enriched components were aged to allow transformation of a proportion of added metals to non-labile forms. Results from the isotopic dilution L value method showed that despite taking up more metals, T. caerulescens accessed the same pool of metals as B. napus. Hence differential access to different solid-phase pools of metals appears to be an unlikely mechanism underlying metal hyperaccumulation. For all components except charcoal, L values for Cd and Zn were greater than the corresponding E values suggesting that E values may tend to underestimate the bioavailable fraction of metals in soils.

Changes in soil biological activities under reduced soil pH during Thlaspi caerulescens phytoextraction

Phytoextraction of soil Cd and Zn may require reduction in soil pH in order to achieve high metal uptake. Reducing the pH of high metal soil, however, could negatively affect soil ecosystem function and health. The objectives of this study were to characterize the quantitative causal relationship between pH and soil biological activities in two Zn and Cd contaminated soils and to investigate the relationship between metals and soil biological activities under low pH. Soils were adjusted to five or six different pH levels by sulfur addition, followed by salt leaching. Thlaspi caerulescens was grown for 6 months, and both the rhizosphere and non-rhizosphere soil biological activities were tested after harvest. Reducing pH significantly lowered soil alkaline phosphatase activity, arylsulphatase activity, nitrification potential, and respiration. However, acid phosphatase activity was increased with decreasing pH. The relationship between soil biological activities and pH was well characterized by linear or quadratic regression models with R 2 values ranging from 0.57 to 0.99. In general, the three enzyme activities, nitrification potential, and the ratio of alkaline phosphatase to acid phosphatase activity were very sensitive indicators of soil pH status while soil respiration was not sensitive to pH change. The rhizosphere soil had higher biological activities than non-rhizosphere soil. The negative effects observed in the non-rhizosphere soil were alleviated by the rhizosphere influence. However, rhizosphere soil after 6 months phytoextraction showed lower nitrification potential than non-rhizosphere soil, probably due to substrate limitation in our study.