Alan J. M. Baker

Accumulators and excluders ‐strategies in the response of plants to heavy metals

Abstract Plants colonizing metalliferous soils have evolved physiological mechanisms which enable them to tolerate metal toxicity. These mechanisms do not generally suppress metal uptake but result in internal detoxification. Two basic strategies of plant response are suggested, accumulators and excluders. In the former, metals can be concentrated in plant parts from low or high background levels. By contrast, differential uptake and transport between root and shoot in excluders, lead to more or less constant low shoot levels over a wide range of external concentration. ‘Indicators’ are seen as a further mode of response where proportional relationships exist between metal levels in the soil, uptake and accumulation in plant parts. The physiological properties of accumulator and excluder species are considered in relation to metal tolerance mechanisms.

Phytoremediation of soil metals

The phytoremediation of metal-contaminated soils offers a low-cost method for soil remediation and some extracted metals may be recycled for value. Both the phytoextraction of metals and the phytovolatilization of Se or Hg by plants offer great promise for commercial development. Natural metal hyperaccumulator phenotype is much more important than high-yield ability when using plants to remove metals from contaminated soils. The hypertolerance of metals is the key plant characteristic required for hyperaccumulation; vacuolar compartmentalization appears to be the source of hypertolerance of natural hyperaccumulator plants. Alternatively, soil Pb and Cr6+ may be inactivated in the soil by plants and soil amendments (phytostabilization). Little molecular understanding of plant activities critical to phytoremediation has been achieved, but recent progress in characterizing Fe, Cd and Zn uptake by Arabidopsis and yeast mutants indicates strategies for developing transgenic improved phytoremediation cultivars for commercial use.

The possibility of in situ heavy metal decontamination of polluted soils using crops of metal-accumulating plants

The decontamination of soils and wastes polluted with heavy metals presents one of the most intractable problems for soil clean-up. Present technology relies upon metal extraction or immobilization processes, both of which are expensive and which remove all biological activity in the soil during decontamination. They may only be appropriate for small areas of valuable redevelopment land. In this paper the use of metal-accumulating plants is explored for the removal of metals from superficially-contaminated soils such as those resulting from the long-term application to land of metal-contaminated sewage sludges. Green remediation employs plants native to metalliferous soils with a capacity to bioaccumulate metals such as zinc and nickel to concentrations greater than 2% in the aerial plant dry matter (hyperaccumulators). Growing such plants under intensive crop conditions and harvesting the dry matter is proposed as a possible method of metal removal and for ‘polishing’ contaminated agricultural soils down to metal concentrations below statutory limits. Not only are the biological activity and physical structure of soils maintained but the technique is potentially cheap, visually unobtrusive and offers the possibility of biorecovery of metals. The limitations of the process are reviewed and the future requirements for the development of efficient phytoremediators are outlined.

Current approaches to the revegetation and reclamation of metalliferous mine wastes

Abandoned metalliferous mine wastes can result in severe pollution and have aesthetic impacts on the local environment. Use of a vegetation cover gives a cost-effective and environmentally sustainable method of stabilising and reclaiming wastes such as mine-spoils and tailings. Many characteristics of metalliferous wastes are often inimical to successful vegetation establishment, most notably phytotoxic levels of residual heavy metals, low nutrient status and poor physical structure of the substratum. Current approaches to revegetation and reclamation involve both ameliorative and adaptive strategies to allow plant establishment and encourage subsequent vegetation development. Different techniques of revegetation are available for temperate and arid, subtropical regions depending on the characteristics of the waste. These include direct seeding with commercially available plants, use of cover and barrier systems and the enhancement of natural revegetation processes.

Hyperaccumulators of metal and metalloid trace elements: Facts and fiction

BackgroundPlants that accumulate metal and metalloid trace elements to extraordinarily high concentrations in their living biomass have inspired much research worldwide during the last decades. Hyperaccumulators have been recorded and experimentally confirmed for elements such as nickel, zinc, cadmium, manganese, arsenic and selenium. However, to date, hyperaccumulation of lead, copper, cobalt, chromium and thallium remain largely unconfirmed. Recent uses of the term in relation to rare-earth elements require critical evaluation.ScopeSince the mid-1970s the term ‘hyperaccumulator’ has been used millions of times by thousands of people, with varying degrees of precision, aptness and understanding that have not always corresponded with the views of the originators of the terminology and of the present authors. There is therefore a need to clarify the circumstances in which the term ‘hyperaccumulator’ is appropriate and to set out the conditions that should be met when the terms are used. We outline here the main considerations for establishing metal or metalloid hyperaccumulation status of plants, (re)define some of the terminology and note potential pitfalls.ConclusionsUnambiguous communication will require the international scientific community to adopt standard terminology and methods for confirming the reliability of analytical data in relation to metal and metalloid hyperaccumulators.

Physiological Characterization of Root Zn2+ Absorption and Translocation to Shoots in Zn Hyperaccumulator and Nonaccumulator Species of Thlaspi.

Radiotracer techniques were employed to characterize 65Zn2+ influx into the root symplasm and translocation to the shoot in Thlaspi caerulescens, a Zn hyperaccumulator, and Thlaspi arvense, a nonaccumulator. A protocol was developed that allowed us to quantify unidirectional 65Zn2+ influx across the root-cell plasma membrane (20 min of radioactive uptake followed by 15 min of desorption in a 100 [mu]M ZnCl2 + 5 mM CaCl2 solution). Concentration-dependent Zn2+ influx in both Thlaspi species yielded nonsaturating kinetic curves that could be resolved into linear and saturable components. The linear kinetic component was shown to be cell-wall-bound Zn2+ remaining in the root after desorption, and the saturable component was due to Zn2+ influx across the root-cell plasma membrane. This saturable component followed Michaelis-Menten kinetics, with similar apparent Michaelis constant values for T. caerulescens and T. arvense (8 and 6 [mu]M, respectively). However, the maximum initial velocity for Zn2+ influx in T. caerulescens root cells was 4.5-fold higher than for T. arvense, indicating that enhanced absorption into the root is one of the mechanisms involved in Zn hyperaccumulation. After 96 h 10-fold more 65Zn was translocated to the shoot of T. caerulescens compared with T. arvense. This indicates that transport sites other than entry into the root symplasm are also stimulated in T. caerulescens. We suggest that although increased root Zn2+ influx is a significant component, transport across the plasma membrane and tonoplast of leaf cells must also be critical sites for Zn hyperaccumulation in T. caerulescens.

Metal ion ligands in hyperaccumulating plants

Metal-hyperaccumulating plants have the ability to take up extraordinary quantities of certain metal ions without succumbing to toxic effects. Most hyperaccumulators select for particular metals but the mechanisms of selection are not understood at the molecular level. While there are many metal-binding biomolecules, this review focuses only on ligands that have been reported to play a role in sequestering, transporting or storing the accumulated metal. These include citrate, histidine and the phytosiderophores. The metal detoxification role of metallothioneins and phytochelatins in plants is also discussed.

Zinc and Cadmium Uptake by Hyperaccumulator Thlaspi caerulescens and Metal Tolerant Silene vulgaris Grown on Sludge-Amended Soils

Two metal tolerant plants, Thlaspi caerulescens J. and C. Presl. (hyperaccumulator), and Silene vulgaris L. (indicator) were grown with Paris Island Cos Romaine lettuce (Lactuca sativa var. longifolia) on longterm sewage sludge plots. Metal uptake patterns by plants in relation to total soil metal and soil pH were examined. The 2-year study used four treatments and two pH levels. Zinc and Cd uptake were measured. Zinc and Cd for Silene and lettuce were as expected with increasing plant concentration in the more contaminated treatments and lower pH levels. Thlaspi followed the same pattern for Cd but not for Zn. Concentrations of Cd were not significantly different between Thlaspi and the other plants. Zinc concentrations in Thlaspi (2000 and 4000 mg kg -1 ) were 10-fold greater than in Silene. They showed no relation to available soil Zn. Although Thlaspi appears to hyperaccumulate Zn on mildly contaminated soils, Cd uptake follows predictable patterns.

Development of a technology for commercial phytoextraction of nickel: economic and technical considerations

In recent R&D work, we have made progress in developing a commercial technology using hyperaccumulator plant species to phytoextract nickel (Ni) from contaminated and/or Ni-rich soils. An on-going program is being carried out to develop a genetically improved phytoextraction plant that combines favorable agronomic and Ni accumulation characteristics. Genetically diverse Ni hyperaccumulator species and ecotypes of Alyssum were collected and then evaluated in both greenhouse and field using serpentine and Ni-refinery contaminated soils. Large genetic variation was found in those studies. Mean shoot Ni concentrations in field-grown plants ranged from 4200 to 20 400 mg kg−1. We have been studying several soil management practices that may affect the efficiency of Ni phytoextraction. Soil pH is an important factor affecting absorption of metals by plants. An unexpected result of both greenhouse and field experiments was that Ni uptake by two Alyssum species was reduced at lower soil pH and increased at higher soil pH. At higher pH, plant yield was improved also. In soil fertility management studies, we found that N application significantly increased plant biomass, but did not affect plant shoot Ni concentration. These findings indicate that soil management will be important for commercial phytoextraction. A number of field trials have been carried out to study planting methods, population density, weed control practices, harvest schedule and methods, pollination control, and seed processing. Such crop management studies have improved phytoextraction efficiency and provide a tool for farmers to conduct commercial production. We have done some work to develop efficient and cost-effective methods of Ni recovery. Recovery of energy by biomass burning or pyrolysis could help make phytoextraction more cost-effective. The progress made in our recent studies will enable us to apply this technology commercially in the near future.

Localization of zinc and cadmium in Thlaspi caerulescens (Brassicaceae), a metallophyte that can hyperaccumulate both metals

Summary The compartmentation of Zn and Cd in roots of Thlaspi caerulescens J. & C. Presl, a metallophyte that hyperaccumulates both metals, was investigated by means of EDAX. Two methods for sample preparation, Na 2 S-fixation and freeze-substitution, were compared. Na 2 S-fixation was not suitable for preventing the loss of metal ions from the samples. According to our results from freeze-substituted samples, Cd accumulated mainly in the apoplast and, to a lesser extent, in vacuoles, whereas Zn was principally found in vacuoles and, to a lesser extent, in cell walls.