Biotechnological applications of serpentine soil bacteria for phytoremediation of trace metals

Serpentine or ultramafic soils are produced by weathering and pedogenesis of ultramafic rocks that are characterized by high levels of Ni, Cr, and sometimes Co, but contain low levels of essential nutrients such as N, P, K, and Ca. A number of plant species endemic to serpentine soils are capable of accumulating exceptionally high concentrations of Ni, Zn, and Co. These plants are known as metal “hyperaccumulators.” The function of hyperaccumulation depends not only on the plant, but also on the interaction of the plant roots with rhizosphere microbes and the concentrations of bioavailable metals in the soil. The rhizosphere provides a complex and dynamic microenvironment where microorganisms, in association with roots, form unique communities that have considerable potential for the detoxification of hazardous materials. The rhizosphere bacteria play a significant role on plant growth in serpentine soils by various mechanisms, namely, fixation of atmospheric nitrogen, utilization of 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source, production of siderophores, or production of plant growth regulators (hormones). Further, many microorganisms in serpentine soil are able to solubilize “unavailable” forms of heavy metal–bearing minerals by excreting organic acids. In addition, the metal-resistant serpentine isolates increase the efficiency of phytoextraction directly by enhancing the metal accumulation in plant tissues and indirectly by promoting the shoot and root biomass of hyperaccumulators. Hence, isolation of the indigenous and stress-adapted beneficial bacteria serve as a potential biotechnological tool for inoculation of plants for the successful restoration of metal-contaminated ecosystems. In this study, we highlight the diversity and beneficial features of serpentine bacteria and discuss their potential in phytoremediation of serpentine and anthropogenically metal-contaminated soils.     

Species adaptation in serpentine soils in Lesbos Island (Greece): metal hyperaccumulation and tolerance

Serpentine (ultramafic) soils, containing relatively high nickel and other metal concentrations, present a stressful environment for plant growth but also a preferred substrate for some plants which accumulate nickel in their tissues. In the present study we focused on: (1) the relationships between serpentine soils of Lesbos Island (Greece) and serpentinophilic species in order to test their adaptation to the ‘serpentine syndrome’, and (2) the Ni-hyperaccumulation capacity of Alyssum lesbiacum, a serpentine endemic, Ni-hyperaccumulating species, recorded over all its distribution for the first time. We sampled soil and the most abundant plant species from the four serpentine localities of Lesbos Island. Soil and leaf elemental concentrations were measured across all the sites. Our results confirmed our hypothesis that serpentinophilic species are adapted to elevated heavy metal soil concentrations but restricting heavy metal concentration in their leaves. We demonstrated that different A. lesbiacum populations from Lesbos Island present differences in Ni hyperaccumulation according to soil Ni availability. Our results highlighted the understanding of serpentine ecosystems through an extensive field study in an unexplored area. Alyssum lesbiacum and Thlaspi ochroleucum emerge as two strong Ni hyperaccumulators with the former having a high potential for phytoextraction purposes.     

Natural variation of nickel,zinc and cadmium (hyper)accumulation in facultative serpentinophytes Noccaea kovatsii and N. praecox

Aims

Data on the variability of hyperaccumulation potential of the facultative serpentinophytes Noccaea kovatsii and N. praecox on different geological substrates are scarce. The aim of this study was to assess the accumulation potential of these two species from ultramafic and non-ultramafic substrates, with special emphasis on the hyperaccumulation of Ni, Zn and Cd.

Methods

Samples of plants and corresponding soils were collected from 16 sites covering a wide range of geochemistry. Elemental concentrations were determined in the roots, shoots and the rhizosphere soils. The pH, particle size distribution and content of organic matter were also determined in the soil samples.

Results

All ultramafic accessions of both species hyperaccumulated Ni with high intraspecific variability. Only one accession of N. kovatsii from a schist soil hyperaccumulated Zn, with also a high Cd accumulation. Accumulation and translocation of Ni in both species were much higher in the ultramafic accessions, whereas Zn accumulation and translocation was found in both ultramafic and non-ultramafic accessions.

Conclusions

Ni accumulation and translocation was restricted to ultramafic populations of both species, whereas it is a species-wide trait for Zn. This study provides new and comprehensive information on the natural variation of hyperaccumulation of Ni, Zn and Cd in N. kovatsii and N. praecox.

     

Nickel hyperaccumulation by Thlaspi montanum var. montanum (Brassicaceae): a constitutive trait

Adaptations to particular stresses may occur only in populations experiencing those stresses or may be widespread within a species. Nickel hyperaccumulation is viewed as an adaptation to high-Ni (serpentine) soils, but few studies have determined if hyperaccumulation ability is restricted to populations from high-Ni soils or if it is a constitutive trait found in populations on both high- and low-Ni soils. We compared mineral element concentrations of Thlaspi montanum var. montanum plants grown on normal and high-Ni greenhouse soils to address this question. Seed sources were from four populations (two serpentine, two non-serpentine) in Oregon and northern California, USA. Plants from all populations were able to hyperaccumulate Ni, showing Ni hyperaccumulation to be a constitutive trait in this species. Populations differed in their ability to extract some elements (e.g., Ca, Mg, P) from greenhouse soils. We noted a negative correlation between tissue concentrations of Ni and Zn. We suggest that the ability to hyperaccumulate Ni has adaptive value to populations growing on non- serpentine soil. This adaptive value may be a consequence of metal-based plant defense against herbivores/pathogens, metal- based interference against neighboring plant species, or an efficient nutrient scavenging system. We suggest that the Ni hyperaccumulation ability of T. montanum var. montanum may be an inadvertent consequence of an efficient nutrient (possibly Zn or Ca) uptake system.     

Metal accumulation by the ultramafic flora of Kosovo

The largest serpentine outcrops in Europe occur in the Balkan Peninsula. Kosovo, as a part of this region, hosts an ultramafic area of 487 km² within its territory. This work reports the first systematic biogeochemical survey on the significant and most representative ultramafic massifs of Kosovo. The aim of this study was (i) to detail the geochemical composition of 12 ultramafic pedons obtained from 10 selected sites chosen as representative for Kosovo, (ii) to inventoriate the flora occurring on these sites and (iii) to identify plant species with potential for use in phytostabilization or phytoextraction purposes. Twelve representative pedons from 10 different sites across the country were excavated and 27 horizon samples were collected. Regarding the serpentine flora, a total of 162 plant taxa located at the ultramafic sites were collected. Soils samples were characterized for basic physico-chemical characteristics and both plant and soil samples were analyzed for chemical composition. The serpentine soils samples displayed a vast array of physico-chemical characteristics which reflected the geochemistry of the bedrock, the degree of weathering and the horizon characteristics. However there appeared to be a relationship between edaphic properties and the occurrence of several plant species. Although most of the plants’ species did not show metal concentrations above 1000 mg kg^?1, Odontarrhena muralis (syn. Alyssum murale Waldst. and Kit). and Noccaea ochroleuca (Boiss and Heldr.) F.K.Mey. (syn.Thlaspi ochroleucum), did, thus meeting the criterion of Ni hyperaccumulating plants. Given the aforementioned, the resilience of these plants to both tolerate and accumulate heavy metals may prove useful for phytostabilization,     

Nickel bioavailability in an ultramafic toposequence in the Vosges Mountains (France)

A serpentinised harzburgite outcrop located in the Vosges Mountains hosts a population of the Ni-hyperaccumulator Thlaspi caerulescens J. & C. Presl. A complete study was undertaken to relate the variability of Ni availability along the ultramafic toposequence to pedogenesis, soil mineralogy and functioning with X-Ray Diffraction, Transmission Electron Microscope observations coupled with Isotopic Exchange Kinetics and diethylenetriamine pentaacetic acid extraction of Ni. The soil profiles ranging from Dystric Cambisol to Hypermagnesic Hypereutric Cambisol were distributed unevenly along the toposequence probably due to geochemical variability of the bedrock and also complex quaternary erosion features. The richest soils were characterised by slight mineral weathering leading to Ni, Cr and Fe accumulation in the B horizons whereas the lowest saturated soils had very low-metal contents. Most soil minerals were inherited from the parent materials and there were only few traces of formation of secondary minerals. Primary minerals (e.g. serpentine, chlorite) contained low Ni concentrations (0.2%) whereas neoformed goethite, mainly in the B horizons of the richest soils, contained up to 4.3% Ni. Ni was probably sorbed onto amorphous Fe oxy−hydroxide particles (oxalate extraction) rather than incorporated within the crystal lattice of goethite. Ni availability in the B horizon of Hypereutric Cambisols was extremely high and so was the oxalate extractable Fe. At the toposequence level, there was a high level of Ni availability in the least weathered soils and a very low-availability level in the more intensively weathered soils (strongly acidic pH). Ni availability was unexpectedly positively correlated to pH and was controlled by soil mineralogy and Ni-bearing mineral phases. Ni hyperaccumulation (above 1,000 mg kg⁻¹) by native T. caerulescens was only reached in the Ni-rich soils as a consequence of the local edaphic factors. Ni uptake by T. caerulescens is strongly regulated by Ni availability in soils and therefore related to pedogenesis.     

Evidence of adaptive tolerance to nickel in isolates of Cenococcum geophilum from serpentine soils

Selection for metal-tolerant ecotypes of ectomycorrhizal (ECM) fungi has been reported in instances of metal contamination of soils as a result of human activities. However, no study has yet provided evidence that natural metalliferous soils, such as serpentine soils, can drive the evolution of metal tolerance in ECM fungi. We examined in vitro Ni tolerance in isolates of Cenococcum geophilum from serpentine and non-serpentine soils to assess whether isolates from serpentine soils exhibited patterns consistent with adaptation to elevated levels of Ni, a typical feature of serpentine. A second objective was to investigate the relationship between Ni tolerance and specific growth rates (μ) among isolates to increase our understanding of possible tolerance/growth trade-offs. Isolates from both soil types were screened for Ni tolerance by measuring biomass production in liquid media with increasing Ni concentrations, so that the effective concentration of Ni inhibiting fungal growth by 50% (EC₅₀) could be determined. Isolates of C. geophilum from serpentine soils exhibited significantly higher tolerance to Ni than non-serpentine isolates. The mean Ni EC₅₀ value for serpentine isolates (23.4 μg ml⁻¹) was approximately seven times higher than the estimated value for non-serpentine isolates (3.38 μg ml⁻¹). Although there was still a considerable variation in Ni sensitivity among the isolates, none of the serpentine isolates had EC₅₀ values for Ni within the range found for non-serpentine isolates. We found a negative correlation between EC₅₀ and μ values among isolates (r = −0.555). This trend, albeit only marginally significant (P = 0.06), indicates a potential trade-off between tolerance and growth, in agreement with selection against Ni tolerance in “normal” habitats. Overall, these results suggest that Ni tolerance arose among serpentine isolates of C. geophilum as an adaptive response to Ni exposure in serpentine soils. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The flora and biogeochemistry of the ultramafic soils of Goiás state, Brazil

Major collections of the ultramafic flora of Goiás, central Brazil, were made by Brooks and co-workers in 1988 and 1990. At the time of reports on this material in 1990–1992 much of it had been identified only tentatively and incompletely, but the area was clearly interesting for taxonomic and biogeochemical reasons. Further progress has been made but still only two-thirds of the specimens are identified at the species level. Following a third collection in early 2005, we now have 800 specimens from this area, with chemical analyses of all the plants and of more than 120 representative soil samples. New species have been found, e.g., in Paspalum (Poaceae) and Pterolepis (Melastomataceae). There is a need for more taxonomic work in genera such as Cnidoscolus (Euphorbiaceae), Lippia (Verbenaceae), Turnera and Piriqueta (Turneraceae), and Vellozia (Velloziaceae). Ni hyperaccumulation (>1,000 mg/kg in dry plant matter) has now been found in a total of 79 specimens, representing more than 30 different species. Notable Ni hyperaccumulators include Pfaffia sarcophylla (Amaranthaceae), species of Justicia, Lophostachys and Ruellia (Acanthaceae), Porophyllum (Asteraceae), several species of Lippia (Verbenaceae), Turnera and Piriqueta (Turneraceae), and a possibly new Cnidoscolus (Euphorbiaceae). Ni hyperaccumulation has not been found in plants of the outcrops of Morro Feio or Crominia-Mairipotaba; it seems to be confined to the extensive layered ultramafics of Barro Alto and the Macedo-Niquelandia areas. The distribution of Ni-values in the Brazilian plant collection is different from that found in the Mediterranean and California, where there is a clear distinction between accumulator and non-accumulator plants: in Brazil the distribution is more continuous, and median Ni concentrations are much greater. An ultramafic hill just north of Niquelandia deserves to be protected because of the presence there of many of the hyperaccumulators and species probably endemic to the Goiás ultramafics.     

Dolomite flora of the Baetic Ranges glades (South Spain)

The vegetation growing on special types of rocks has long attracted the attention of botanists and ecologists. In this respect, interest in serpentine soils has led to the publication of a number of monographs on their flora and vegetation. The high levels of heavy metals, in particular of Ni, and the low proportion of Ca/Mg are generally held responsible for the severe restrictions limiting flora development on this kind of rocks. Although researchers have also studied other types of unusual bedrocks such as gypsum outcrops, little work has been done on dolomites. This is surprising because dolomites share certain features with serpentines (they are both rocks rich in Mg). This paper deals with the flora peculiar to the dolomites of the Baetic Ranges, one of the richest territories in Europe as far as flora is concerned. A number of botanists with experience in the study of this very particular flora have joined together to make a list of the plants peculiar to the dolomites. Our catalogue includes 144 species with a variable degree of “dolomitophily”, the score of which ranging from 1 to 3, the maximal value. A comparison of the taxonomical spectrum of this catalogue with the one found in territories where dolomites are dominant or with other areas of serpentines reveals that there is a given deviation or bias in favour of some families. A similar comparison between biotypes also leads to the observation that, as usually happens on ultramafic rocks, hemicryptophytes tend to be dominant on dolomites. In addition, we show that the proportion of Ca/Mg catalogued for different kinds of soils could be a key element in explaining the restrictions which dolomites and serpentines impose on plants. In dolomites this proportion shows an intermediate position (2.19) between soils derived from limestones (11.30) and serpentines (0.84) in the study area.     

Hyperaccumulation of nickel by two Alyssum species from the serpentine soils of Iran

Serpentine soils, which contain relatively high concentrations of nickel and some other metals, are the preferred substrate for some plants, especially those that accumulate Ni in their tissues. In temperate regions more Ni-hyperaccumulator plants are found in Alyssum than in any other genus. In this study, serpentine soils of two areas (Marivan and Dizaj) in the west/northwest of Iran and also perennial Alyssum plants growing on these soils were analyzed for Ni and some other metals. The highest concentrations of total metals in the soils of these areas for Ni, Cr, Co and Mn were 1,350, 265, 94 and 1,150 μg g⁻¹, respectively, while concentrations of Fe, Mg and Ca reached 3.55%, 16.8% and 0.585% respectively. The concentration of exchangeable Ni in these soils is up to 4.5 μg g⁻¹. In this study two Alyssum species, A. inflatum and A. longistylum, have been collected from Marivan and Dizaj, respectively. Analysis of leaf dry matter shows that they can contain up to 3,700 and 8,100 μg Ni g⁻¹, respectively. This is the first time that such high Ni concentrations have been found in these species. The concentrations of other metals determined in these species were in the normal range for serpentine plants, except for Ca, which was higher, up to 5.3% and 3.5%, respectively     

Tolerance and induction of tolerance to Ni of arbuscular mycorrhizal fungi from New Caledonian ultramafic soils

The influence of Ni on arbuscular mycorrhizal fungi (AMF) has not been studied yet. We tested the tolerance to Ni of five AMF isolates from New Caledonian ultramafic soils. Spore germination indicated that these isolates were clearly more tolerant to Ni than three other isolates from non-ultramafic soils. They were able to germinate at 30 μg g⁻¹ Ni, whereas spores of the non-ultramafic isolates were totally inhibited at 15 μg g⁻¹ Ni. Among the ultramafic isolates, two were obtained from roots of Ni-hyperaccumulating plants. Their tolerance to Ni was clearly higher than all the other isolates. The proportion of germinated spores of the different isolates in contact with ultramafic soils showed the same tendencies as those observed with Ni solutions. Tolerance to Ni increased when spores were produced from mycorrhiza on plants grown on sand containing 20 μg g⁻¹ Ni, in comparison with those produced on sand without Ni. These results indicate that the tolerance to Ni of AMF spores can be induced by the presence of this metal in the substrate.

Richness and diversity of Pontic flora on serpentine of Serbia

Serpentine substrate in Serbia covers wide areas mainly in Kosovo and in central, western and south-western part of the country. On the serpentines of Serbia, particularly on the skeleton soils of hills and on lower elevation mountain slopes, there are some xerophilous and steppe-like vegetations. These xerophilous grasslands include mostly Pontic, Pontic-Submediterranean and/or Mediterranean-Submediterranean-Pontic plants. The distribution of 161 taxa of Pontic, Pontic-Submediterranean and Mediterranean-Submediterranean-Pontic areal — types on serpentinites of Serbia was analyzed in order to distinguish centers of richness and diversity. The distribution of taxa was analyzed with respect to geographic, ecological and climatic factors. The qualitative composition of Pontic flora on the serpentine areas was also compared. The greatest floristic richness and the center of diversity of Pontic flora on serpentine of Serbia was recorded in central part of the area investigated.     

Distribution and Metal-Accumulating Behavior of Thlaspi caerulescens and Associated Metallophytes in France

The heavy metal hyperaccumulator Thlaspi caerulescens is widespread in France on many kinds of sites and substrates, including Zn/Pb/Cd mine and smelter wastes, Ni-rich serpentine outcrops and a variety of nonmetalliferous soils. Thlaspi caerulescens is remarkable among the metallophytes of France because it accumulates Zn to high concentrations (almost always >0.1%, and often >1% in the dry matter) regardless of the total Zn concentration of the substrate. The extraordinary uptake of Zn from soils of normal Zn concentration draws attention to the need for studies of the mechanisms by which such mobilization and uptake can occur. Different populations of Thlaspi caerulescens in France show considerable variation in their ability to accumulate Cd; individuals in some populations contain as much as 0.1 to 0.4% Cd, the highest levels recorded in vascular plants. The hyperaccumulation of Ni (sometimes exceeding 1%) from serpentine soils in France is also noteworthy. Despite the generally low biomass, some very large individuals occur, giving good potential for selective breeding to improve the value of Thlaspi caerulescens for phytoremediation, especially of Cd. The high Zn uptake from all kinds of soils is a property shared by the related T. brachypetalum, and T. alpinum shows dual Zn- and Ni uptake, depending on the substrate. The extent to which other species of Thlaspi occurring in France exhibit metal accumulation is also discussed.     

Within and between population variation for zinc and nickel accumulation in two species of Thlaspi (Brassicaceae)

In this study, the differences in zinc (Zn) and nickel (Ni) hyperaccumulation were investigated between three populations of Thlaspi pindicum together with genetic variation within populations of T. pindicum and Thlaspi alpinum var. sylvium, both serpentine endemics. Three experiments were conducted under standard conditions in hydroponic assay. Each experiment contained three treatments of metal: 100 microm Zn, 100 microm Ni, and combined 100/100 microm Zn/Ni. Genetic variation within populations was determined using maternal families. No genetic variation within populations was found for either Zn or Ni hyperaccumulation for both T. pindicum and T. alpinum var. sylvium, but differences were observed for both Zn and Ni hyperaccumulation between populations of T. pindicum. In combined Zn/Ni treatments, Zn inhibited Ni translocation in both species, which is unexpected considering that these species are serpentine endemics and well known Ni hyperaccumulators. The lack of genetic variation for metal hyperaccumulation is possibly due to inbreeding. Since Zn hyperaccumulation is not manifested in the field, inadvertent uptake of Zn is a plausible hypothesis for its preferential uptake.     

Responses of two populations of an Iranian nickel-hyperaccumulating serpentine plant,Alyssum inflatum Nyar., to substrate Ca/Mg quotient and nickel

We selected two geographically close serpentine and non-serpentine populations of a Ni-hyperaccumulating plant (Alyssum inflatum) to investigate the influence of two common factors of serpentine soils: high Ni concentrations and low Ca/Mg quotients. Soils and plants were sampled from serpentine and non-serpentine substrates, and concentrations of Ca, Mg and Ni were measured. A hydroponic culture was used to compare growth and elemental composition responses of serpentine and non-serpentine plants to different Ca/Mg quotients and Ni concentrations in the nutrient solution. The Ca/Mg quotient for non-serpentine soils was 15 times higher than for serpentine soils, but there was no difference in the Ca/Mg quotient of plants from the two populations. In hydroponic culture, plants from both populations were able to survive at high Ca/Mg quotients. This result suggests that serpentine plants of A. inflatum do not necessarily need a substrate with a low Ca/Mg quotient for survival. Decreases in the Ca/Mg quotient in hydroponics decreased growth. The magnitude of this decrease was significantly greater in non-serpentine plants, suggesting a greater resistance of serpentine plants to low Ca/Mg quotients. Total Ni concentration in serpentine soils was 13 times higher than in non-serpentine soils, but ammonium nitrate-extractable concentrations of Ni in both soil types were similar. Ni concentrations in non-serpentine plants from their natural habitat were significantly lower than in serpentine plants, but there was no significant difference in Ni accumulation by plants of the two populations in hydroponic culture. However, increased concentrations of Ni in the hydroponic medium caused similar decreases in growth of both populations, indicating that Ni tolerance of the two populations was similar.     

The ecological significance of nickel hyperaccumulation: a plant chemical defense

Nickel hyperaccumulating plants have more than 1000 mg Ni kg^–1 dry weight when grown on nickel-bearing soils. We hypothesized that Ni hyperaccumulation could serve as a chemical defense against herbivores In feeding experiments with potential insect herbivores and Ni hyperaccumulating plants, only those inseets fed leaves from plants grown on non-nickel-bearing soil survived or showed a weight gain. Among chemical parameters measured, only Ni content of plants was sufficient to explain this result. When subjected to herbivory by lepidopteran larvae, plants grown on Ni-amended soil showed greater survival and yield than plants on unamended soil. Ni hyperaccumulation may be an effective plant chemical defense against herbivores because of its high lethality, apparent low cost, and broad spectrum of toxicity.     

A role for nickel in osmotic adjustment in drought-stressed plants of the nickel hyperaccumulator <Emphasis Type="Italic">Stackhousia tryonii</Emphasis> Bailey

The hypothesis that hyperaccumulation of certain metals in plants may play a role in osmotic adjustment under water stress (drought) was tested in the context of nickel hyperaccumulator Stackhousia tryonii. Field-collected mature plants of S. tryonii, grown in native ultramafic soil, were pruned to soil level and the re-growth exposed to five levels of water stress (20, 40, 60, 80 and 100% field capacity; FC) for 20 weeks. Water stress had significant (P<0.05) influence on growth (biomass), water potential and shoot Ni concentrations, with progressively more impact as water stress was increased from 80 to 40% FC. Shoot Ni concentration increased significantly from 3,400 μg g⁻¹ dry weight (at 100% FC) to 9,400 μg g⁻¹ dry weight (at 20% FC). Assuming that Ni is uniformly distributed through the shoot tissue, the Ni concentration could account for 100% at the 80 and 60% FC conditions, and 50% at the 40 and 20% FC conditions of plant osmotic regulation. The results are consistent with a role of Ni in osmotic adjustment and protection of S. tryonii plants against drought.     

A phytogeochemical survey of the flora of ultramafic and adjacent normal soils in North Morocco

Variation in plant elemental composition (Ni, Ca, Mg, Mg/Ca ratio) in relation to soil composition was investigated in a poorly studied ultramafic area in the north of Morocco. A total of 142 leaf samples representing 36 species from 9 sites (5 ultramafic and 4 normal soils from adjacent areas) were analysed. The soil was richer in Mg and Ni and had a higher Mg/Ca ratio in the ultramafic sites than in the control sites, and these differences were qualitatively reflected in the average mineral composition of the plants. However, there were considerable differences in mineral composition among species within serpentinic sites, indicating that species with contrasting mineral nutrition strategies can cope with the mineral element imbalance characteristic of ultramafic soils. Particularly noteworthy was the finding that species with high requirements of Ca are not excluded from serpentinic soils. In view of their high responsiveness to soil nickel and magnesium concentration, Dittrichia viscosa and Lavandula dentata are proposed as bioindicators of these elements in the soil in the Rif area. By contrast, two local serpentine endemics, Halimium atriplicifolium and Notholaena marantae were excluders of nickel and magnesium. This revised version was published online in August 2006 with corrections to the Cover Date.     

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.     

Genetic Diversity of Bacterial Communities of Serpentine Soil and of Rhizosphere of the Nickel-Hyperaccumulator Plant <Emphasis Type="Italic">Alyssum bertolonii</Emphasis>

Serpentine soils are characterized by high levels of heavy metals (Ni, Co, Cr), and low levels of important plant nutrients (P, Ca, N). Because of these inhospitable edaphic conditions, serpentine soils are typically home to a very specialized flora including endemic species as the nickel hyperaccumulator Alyssum bertolonii. Although much is known about the serpentine flora, few researches have investigated the bacterial communities of serpentine areas. In the present study bacterial communities were sampled at various distances from A. bertolonii roots in three different serpentine areas and their genetic diversity was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis. The obtained results indicated the occurrence of a high genetic diversity and heterogeneity of the bacterial communities present in the different serpentine areas. Moreover, TRFs (terminal restriction fragments) common to all the investigated A. bertolonii rhizosphere samples were found. A new cloning strategy was applied to 27 TRFs that were sequenced and taxonomically interpreted as mainly belonging to Gram-positive and -Proteobacteria representatives. In particular, cloned TRFs which discriminated between rhizosphere and soil samples were mainly interpreted as belonging to Proteobacteria representatives.This revised version was published online in November 2004 with corrections to Volume 48.