The potential of Thlaspi caerulescens for phytoremediation of contaminated soils

Uptake of Cd, Zn, Pb and Mn by the hyperaccumulator Thlaspi caerulescens was studied by pot trials in plant growth units and in populations of wild plants growing over Pb/Zn base-metal mine wastes at Les Malines in the south of France. The pot trials utilised metal-contaminated soils from Auby in the Lille area. Zinc and Cd concentrations in wild plants averaged 1.16% and 0.16% (dry weight) respectively. The unfertilised biomass of the plants was 2.6 t/ha. A single fertilised crop with the above metal content could remove 60 kg of Zn and 8.4 kg Cd per hectare. Experiments with pot-grown and wild plants showed that metal concentrations (dry weight basis) were up to 1% Zn (4% Zn in the soil) and just over 0.1% Cd (0.02% Cd in the soil). The metal content of the plants was correlated strongly with the plant-available fraction in the soils as measured by extraction with ammonium acetate and was inversely correlated with pH. Bioaccumulation coefficients (plant/soil metal concentration quotients) were in general higher for Cd than for Zn except at low metal concentrations in the soil. There was a tendency for these coefficients to increase with decreasing metal concentrations in the soil. It is proposed that phytoremediation using Thlaspi caerulescens would be entirely feasible for low levels of Cd where only a single crop would be needed to halve a Cd content of 10 g/g in the soil. It will never be possible to remediate elevated Zn concentrations within an economic time frame (<10 yr) because of the lower bioaccumulation coefficient for this element coupled with the much higher Zn content of the soils.     

Uptake and transport of zinc in the hyperaccumulator Thlaspi caerulescens and the non-hyperaccumulator Thlaspi ochroleucum

Growth and zinc uptake of the hyperaccumulator species Thlaspi caerulescens J. & C. Presl and the non-hyperaccumulator species Thlaspi ochroleucum Boiss. & Heldr. were compared in solution culture experiments. T. caerulescens was able to tolerate 500 mmol m^?3 (32.5 g m^?3) Zn in solution without growth reduction, and up to 1000 mmol m^?3 (65 g m^?3) Zn without showing visible toxic symptoms but with a 25% decrease in dry matter (DM) yield. Up to 28 g kg^?1 of Zn in shoot DM was obtained in healthy plants of T. caerulescens. In contrast, T. ochroleucum suffered severe phytotoxicity at 500 mmol m^?3 Zn. Marked differences were shown in Zn uptake, distribution and redistribution between the two species. T. caerulescens had much higher concentrations of Zn in the shoots, whereas T. ochroleucum accumulated higher concentrations of Zn in the roots. When an external supply of 500 mmol m^?3 Zn was withheld, 89% of the Zn accumulated previously in the roots of T. caerulescens was transported to the shoots over a 33 d period, whereas in T. ochroleucum only 32% was transported. T. caerulescens was shown to have a greater internal requirement for Zn than other plants. Increasing the supply of Zn from 1 to 10 mmol m^?3 gave a 19% increase in the total DM of this species. liven the shoots from the 1 mmol m^?3 Zn treatment which showed Zn deficiency contained 10 times greater Zn concentrations than the widely reported critical value for Zn deficiency to occur in many other plant species. The results obtained suggest that strongly expressed constitutive sequestration mechanisms exist in the hyperaccumulator T. caerulescens, which detoxify the large amount of Zn present in shoot tissues and decrease its physiological availability in the cytosol. Both T. caerulescens and T. ochroleucum had constitutively high concentrations of malate in shoots, which were little affected by different Zn treatments. Although malate may play a role in Zn chelation because of the high concentrations present, it cannot explain the species specificity of Zn tolerance and hyperaccumulation.     

Phytoextraction of cadmium with Thlaspi caerulescens

The in situ phytoextraction of cadmium from soils can only be achieved using plants that are both tolerant to high Cd concentrations and able to extract sufficient amounts of the metal. However, very few plant species are capable of remediating Cd polluted soils in a reasonable time frame. This paper aims to show that the population of the hyperaccumulator Thlaspi caerulescens J. & C. Presl. from Viviez (south of France), which has a high Cd-accumulating capability, is an efficient tool to remove Cd from contaminated soils. Roots of T. caerulescensViviez proliferate in hot spots of metals in soils which is particularly advantageous because of heterogeneity of the distribution of metal in polluted soils. Isotopic techniques showed that plants from this population acquire Cd from the same pools as non-accumulating species, but that it was much more efficient than non-hyperaccumulators at removing the metal from the soil labile pool. This is due: to (i) a specific rooting strategy, and (ii) a high uptake rate resulting from the existence in this population of Cd-specific transport channels or carriers in the root membrane. Growth and overall extraction can be improved with appropriate N fertilisation, supplied either as mineral fertilisers or uncontaminated sewage sludge. Selecting bigger plants is possible from within a suitable Cd-accumulating population to improve the phytoextraction process. Growing the Cd-accumulating populations results in a reduction in the availability of Cd and Zn as shown with field and lysimeter experiments conducted for several years. As a result, on a practical aspect, Cd hyperaccumulating populations of T. caerulescens may be used as a tool to efficiently reduce the availability of Cd in soils, providing appropriate populations are used.     

Solubility of zinc and interactions between zinc and phosphorus in the hyperaccumulator Thlaspi caerulescens

The relationship between Zn and P in the Zn hyperaccumulator Thlaspi caerulescens J. & C. Presl was investigated using hydroponic culture. Total concentrations of Zn in the shoots increased from 0·2 to 27 g kg^–1 dry mass when solution Zn increased from 1 to 1000 mmol m^–3. Water-soluble Zn accounted for > 80% of the total Zn in the shoots containing > 5 g Zn kg^–1 dry mass. Total P was maintained at about 3 g kg^–1 dry mass in the shoots containing < 20 g Zn kg^–1 dry mass, but significantly decreased with higher Zn concentrations. Linear regression between insoluble P and insoluble Zn in the shoots produced a small slope, suggesting that co-precipitation of Zn and P was not an important detoxification mechanism in the shoots. In contrast, there was a strong correlation between insoluble P and insoluble Zn in the roots, with a linear slope of 0·3 — close to the P:Zn ratio in Zn₃(PO₄)₂. Foliar sprays of phosphate did not affect shoot dry mass significantly, but decreased root length and root dry mass significantly at Zn concentrations in solution from 10 to 3000 mmol m^–3. Foliar P was translocated to roots to enhance co-precipitation of Zn and P, although this did not enhance Zn tolerance. The results suggest that T.caerulescens possesses mechanisms which allow it to accumulate and sequester huge amounts of Zn in the shoots without causing P deficiency.     

Variation in root-to-shoot translocation of cadmium and zinc among different accessions of the hyperaccumulators Thlaspi caerulescens and Thlaspi praecox

Efficient root-to-shoot translocation is a key trait of the zinc/cadmium hyperaccumulators Thlaspi caerulescens and Thlaspi praecox, but the extent of variation among different accessions and the underlying mechanisms remain unclear. Root-to-shoot translocation of Cd and Zn and apoplastic bypass flow were determined in 10 accessions of T. caerulescens and one of T. praecox, using radiolabels (109)Cd and (65)Zn. Two contrasting accessions (Pr and Ga) of T. caerulescens were further characterized for TcHMA4 expression and metal compartmentation in roots. Root-to-shoot translocation of (109)Cd and (65)Zn after 1 d exposure varied 4.4 to 5-fold among the 11 accessions, with a significant correlation between the two metals, but no significant correlation with uptake or the apoplastic bypass flow. The F(2) progeny from a cross between accessions from Prayon, Belgium (Pr) and Ganges, France (Ga) showed a continuous phenotype pattern and transgression. There was no significant difference in the TcHMA4 expression in roots between Pr and Ga. Compartmentation analysis showed a higher percentage of (109)Cd sequestered in the root vacuoles of Ga than Pr, the former being less efficient in translocation than the latter. Substantial natural variation exists in the root-to-shoot translocation of Cd and Zn, and root vacuolar sequestration may be an important factor related to this variation.     

Cadmium accumulation in populations of Thlaspi caerulescens and Thlaspi goesingense

The capacity to accumulate cadmium (Cd) and zinc (Zn) was compared in Thlaspi goesingense and four populations of Thlaspi caerulescens . Two populations of T. caerulescens were grown in hydroponics at five concentrations of Cd. In addition, plants were grown in pots containing compost in which three different concentrations of Cd and two concentrations of Zn were added. A field trial was conducted to compare Zn and Cd uptake by three populations of T. caerulescens on nine selected plots of the Woburn Market Garden Experiment (UK) which had been contaminated to different degrees with heavy metals owing to past applications of sewage sludge. Results show that the four populations of T. caerulescens had the same ability to hyperaccumulate Zn but were significantly different in terms of Cd accumulation. Two populations of T. caerulescens from Southern France accumulated much more Cd than the populations from Prayon (Belgium) and Whitesike (UK). Generally, uptake of Cd was not decreased by increased concentrations of Zn in the substrate. These results indicate that the mechanisms of Cd and Zn hyperaccumulation are not identical in this species. This is the first report of hyperaccumulation of Cd by T. goesingense , but the growth of this species was markedly reduced by the large concentrations of Zn in the substrate. Future work should focus on the differences between Cd and Zn uptake in hyperaccumulator plants at the species and population level.     

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.     

Zinc and cadmium hyperaccumulation by Thlaspi caerulescens from metalliferous and nonmetalliferous sites in the Mediterranean area: implications for phytoremediation

Growth, tolerance and zinc and cadmium hyperaccumulation of Thlaspi caerulescens populations from three metal contaminated soils and three normal soils were compared under controlled conditions. Individuals of six populations were cultivated on five soils with increasing concentrations of zinc (50–25000 μg g⁻¹) and cadmium (1–170 μg g⁻¹). There was no mortality of normal soil populations in the four metal-contaminated soils, but plant growth was reduced to half that of populations from metal-contaminated soils. However, in noncontaminated soil, the growth of individuals from normal soils was greater than that of individuals from metal-contaminated soils. Individuals from normal soils concentrated three times more zinc in the aboveground biomass than those from metal-contaminated soils, but the latter accumulated twice as much cadmium. We conclude that populations of T. caerulescens from both normal and metal-contaminated soils are interesting material for phytoextraction of zinc and cadmium, but to optimize the process of phytoextraction it is necessary to combine the extraction potentials of both type of populations.     

Availability of cadmium and zinc accumulated in the leaves of Thlaspi caerulescens incorporated into soil

When grown on contaminated soil, hyperaccumulator plants contain high concentrations of metals which may return to the soil after senescence. This work was undertaken to assess the availability of Cd and Zn associated to the leaves of the hyperaccumulator Thlaspi caerulescens after incorporation into an uncontaminated soil. A Zn- and Cd- accumulator population of T. caerulescens was grown on a Cd- and Zn- contaminated soil previously labelled with ¹⁰⁹Cd. Leaves (TCL) were harvested, dried, ground and incorporated into the soil at a rate of 2.07 mg Cd kg⁻¹ and 51.9 mg Zn kg⁻¹. Then a pot experiment was conducted for 3 months with rye grass (Lolium perenne) and T. caerulescens. Rye grass was harvested monthly and T. caerulescens at the end of the experiment. Plant biomass was measured, along with the concentration of Cd, Zn and ¹⁰⁹Cd. Results showed that water-extractable metals in TCL were 69% for Zn and 33% for Cd. Addition of TCL to soil, depleted growth of rye grass, and improved that of T. caerulescens. At harvest, concentrations of both metals were increased in plants by TCL. Concentrations of Cd in rye grass increased with the cut number, while that of Zn decreased slightly. Rye grass extracted 1.6% of the total Cd and 0.9% of the total Zn, and T. caerulescens extracted up to 22.4% of the Cd and 7% of the Zn. About 94% of the Cd in rye grass and 86% in T. caerulescens was derived from TCL. In conclusion, metals associated with leaves of the hyperaccumulator T. caerulescens were very mobile after incorporation into the soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

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.     

Heavy metal uptake and chemical changes in the rhizosphere of Thlaspi caerulescens and Thlaspi ochroleucum grown in contaminated soils

Thlaspi caerulescens (J. and C. Presl) and Thlaspi ochroleucum (Boiss. ex Heldr) were grown in three different soils containing moderate to high amounts of heavy metals in a pot experiment, using a rhizobag technique. T. caerulescens accumulated significantly more Zn in the shoots than T. ochroleucum. The concentrations of Zn in the shoots of T. caerulescens ranged from 3100 to 8100 mg kg^-1 dry matter (DM), but only from 800 to 1600 mg kg^-1 DM in T. ochroleucum. Total uptake of Zn in the shoots of T. caerulescens was about 5 times that of T. ochroleucum. In contrast, the differences between the two species in the uptake of Cd, Cr, Cu, Ni and Pb were generally small. Concentrations of mobile Zn (extractable with 1M NH₄NO₃) in the rhizosphere and non-rhizosphere soils decreased considerably after growth of both plants, but the decreases were greater with T. caerulescens than with T. ochroleucum. The decreases in the mobile fraction accounted for less than 10% of the total Zn uptake by T. caerulescens indicating that this species was effective in mobilising Zn from less soluble fractions in the soils. The rhizosphere soils tended to have higher concentrations of mobile Zn than the non-rhizosphere soils, probably because of the lower pH in the rhizosphere. The pH in the rhizosphere soils was 0.2-0.4 units lower than that in the non-rhizosphere soils at the end of the experiment. However, there were no significant differences between the two species in the degree of rhizosphere acidification. The results suggest that T. caerulescens has potential for removing Zn from moderately to highly contaminated soils, but that this ability was not related to the pH changes in the rhizosphere.     

Palatability of Thlaspi caerulescens for snails: influence of zinc and glucosinolates

* The hypothesis that zinc (Zn) hyperaccumulation defends Thlaspi caerulescens against herbivores is tested with the snail Helix aspersa. We investigated the effects of leaf zinc, cadmium, glucosinolate, nitrogen and dry matter concentrations on the feeding preferences of snails. * Four T. caerulescens populations from southern France (two from metalliferous and two from normal soils) were grown on low- and high-Zn soils to obtain contrasting leaf Zn concentrations. Plants were also collected in the field, and binary feeding choices involving low- and high-Zn leaves were conducted. * Foliar Zn, Cd, N and dry matter concentrations did not affect the feeding choices of snails, whereas glucosinolate had a significant negative effect on herbivore preferences. Compared with metallicolous plants, nonmetallicolous ones appeared to be better protected against snails, whatever their Zn concentration. * These results do not support the defence hypothesis, as glucosinolates appear to decrease the degree of herbivory when Zn does not.     

Intraspecific variation of metal preference patterns for hyperaccumulation in Thlaspi caerulescens: evidence from binary metal exposures

Metal preferences with regard to accumulation were compared between populations of the heavy metal hyperaccumulator Thlaspi caerulescens, originating from calamine, serpentine and non-metalliferous soils. Plants were exposed for 3 weeks to factorial combinations of concentrations of different metals in binary mixture in hydroponics. The nature and degree of the interactions varied significantly between populations. In the calamine, non-Cd/Ni-hyperaccumulating population, La Calamine, there were no one-sided or mutual antagonistic interactions among the metals with regard to their accumulation in the plant. In three other populations capable of Cd and Ni hyperaccumulation, from calamine, serpentine and non-metalliferous soil respectively, there were one-sided or mutual antagonistic interactions between Cd and Zn, Cd and Ni, and Ni and Zn, possibly resulting from competition for transporters involved in uptake or plant-internal transport. Significant synergistic interactions, probably resulting from regulation of transporter expression, were also found, particularly in the La Calamine population. All the populations seemed to express a more or less Zn-specific high-affinity system. The serpentine and the non-metallicolous populations seemed to posses low-affinity systems with a preference for Cd and Zn over Ni, one of which may be responsible for the Ni hyperaccumulation of the serpentine population in its natural environment. The calamine population from Ganges also seemed to express a strongly Cd-specific high-affinity system which is in part responsible for the Cd-hyperaccumulation phenotype exhibited by this population in its natural environment.