Heavy Metal Adaptation Secondary article

Luis Rafael Herrera-Estrella, CINVESTAV of Irapuato, Irapuato, Guanajuato, Mexico Article Contents
Angel Arturo Guevara-Garcı́a, CINVESTAV of Irapuato, Irapuato, Guanajuato, Mexico . Introduction
. Differences in Ecology
José López-Bucio, CINVESTAV of Irapuato, Irapuato, Guanajuato, Mexico
. Colonizers of Specific Sites
. Mechanisms of Metal Uptake, Exclusion, Transport and
The adaptive responses of living organisms to heavy metals include physiological, Storage
anatomical and ecological mechanisms, some of which are highly conserved in nature.
These adaptive mechanisms enable certain species to survive even in the presence of high
concentrations of these toxic elements.

Introduction and grow in the presence of potentially toxic concentra-

Heavy metals are defined as the group of elements that
have densities higher than 5 g cm 2 3. Approximately 40
chemical elements fall in this category. All heavy metals are
toxic at high concentrations, including those that are
essential micronutrients, such as copper and molybdenum.
Therefore, living species must actively respond to protect
themselves from metal poisoning in contaminated sites.
Heavy metals are mainly localized as disperse compo-
nents in rock formations and few ecosystems present
natural heavy metal toxicity, such as aluminium toxicity in
acid soils. However, in a considerable number of places,
heavy metal concentrations in soil increase to toxic levels
through agricultural, manufacturing, mining and waste
disposal practices. Recent findings suggest that the
adaptive response of living organisms to contaminated
environments can be a rapid and efficient process. Insights
into the mechanisms involved in heavy metal adaptation
are beginning to be obtained.
Differences in Ecology
Soil properties and the adaptive response
The distribution of living organisms in ecosystems affected
by heavy metal toxicity depends not only on the kind,
combination and concentration of metals, but also on
environmental factors such as soil chemistry, heterogeneity
and, in particular, nutrient status (principally nitrogen,
phosphorus and potassium).
Gradients of soil conditions often determine areas of
increasing stress or disturbance that impose a high selective
pressure on the colonizing species. During evolution,
adaptations have developed in a very wide range of
organisms from all major taxonomic groups. This is
reflected in the high number of species of some taxa that
have become endemic in sites polluted with heavy metals.
In each case, these organisms have the ability to survive
tions of heavy metals.
Heavy metal effects in natural populations
Toxic metals are believed to affect natural populations by
reducing the abundance and diversity of species and
selecting for resistant/tolerant populations. Several studies
have shown the high specificity of species distribution in
metal-poisoned ecosystems. In fungi, the reduction in
number of species has been noted in soils polluted with
copper, cadmium, lead, arsenic and zinc. In plants, the
formation of endemisms has been documented in different
ecosystems around the world, for example in the tropical
island of New Caledonia (around 16 000 km2), where
serpentine soils containing toxic levels of nickel, chromium
and cobalt cover about one third of the island surface.
These typically infertile soils contain a native flora of
approximately 1500 species comprising woody perennials,
epiphytic orchids and Cyperaceae growing in xerophytic
scrub at low altitude (Morat et al., 1986). In the copper
region of Upper Shaba in Zaire, which comprises about
100 copper–cobalt ore deposits totalling some 20 km2,
disseminated in a metallogenic province, a number of
distinct metalliferous habitats exist, such as copper
clearings and natural isolated copper hillocks. These bear
specific plants, notably small annual herbs and grasses.
About 220 taxa are present on these outcrops, of which 42
are endemic. Some of these are located on a single hillock
(Malaisse, 1983). In the mining area of Stolberg (South
Aachen, Germany), Betula trees grow well on hills of
mining smelter ash with a total lead content of up to 10–
20 g per kg. In each of these cases, the diversity of plant
species is severely restricted, and often edaphic ecotypes,
which are tolerant to the specific metal present in excess in
the soil, are selected rapidly. Endemisms are typical in
areas polluted for fewer than 20 years. At roadsides and
beneath galvanized fences are habitats where adaptation
takes place rapidly.

ENCYCLOPEDIA OF LIFE SCIENCES © 2001, John Wiley & Sons, Ltd. www.els.net 1
 Heavy Metal Adaptation
Symbiosis events in contaminated
environments
Ecological associations can be found along polluted
habitats. There are some examples that suggest a causal
role in the reduction of metal phytotoxicity by mycorrhizal
fungi. In ericaceous plants, little or no growth occurs in
mycorrhiza-free plants in the presence of copper and zinc;
however, two species of ectomycorrhizal fungi, Amanita
muscaria and Paxillus involutus, increased zinc tolerance of
Betula sp. From these examples, it was suggested that the
hyphal complexes of the mycorrhizal fungi bind metals,
thus preventing metal translocation to the plant and the
resulting toxic symptoms (Bradley et al., 1982).
Colonizers of Specific Sites
The ability of certain bacterial, fungal and plant species to
colonize environments polluted by heavy metals has been
widely described. Metal-tolerant anaerobic bacteria iso-
lated from contaminated habitats use toxic metals (such as
selenium and chromium) as electron acceptors. The
bacterial genera Thiobacillus, Streptomyces, Streptococcus
and Caulobacter have been isolated from sites polluted by
mercury ions. In these cases, the mechanism of detoxifica-
tion seems to involve the intracellular reduction of
toxic forms (Hg2 1 ) to the less toxic, relatively inert
metallic form (Hg0) by the activity of specific enzymes
(mercury reductases). The accumulation of cadmium in
Citrobacter and uranium in strains of Pseudomonas has
also been found.
In the river Rio Tinto in Spain, which has an
exceptionally low pH (around two) and high concentra-
tions of heavy metals (such as iron, arsenic, copper,
cadmium and nickel), almost 1300 species have been
collected. The bacterium Thiobacillus ferrooxidans and
Leptospirillum ferrooxidans are especially abundant and,
surprisingly, patches of algae and masses of filamentous
fungi living together with several species of yeast and
protist have also been found (Ariza, 1998).
In soils with highly toxic levels of copper and zinc, the
fungi Geomyces and Paecilomyces have been found to be
the predominant genera, whereas Penicillium and Oidio-
dendron spp. decline significantly. In samples taken from
an organomercurial-treated golf green, the species Tricho-
cladium asperum, Trichoderma hamatum, Zygorrhynchus
moelleri and Chrysosporium pannorum have been found
frequently, whereas the genera Chaetomium, Fusarium,
Penicillium and Paecilomyces are greatly reduced. In soil
contaminated with cadmium dust, Strobilurus tenacellus,
Mycena ammoniaca and Armillaria lutea are the most
common basidiomycetes.
Some Penicillium species are sensitive towards heavy
metals, but one of the best examples of fungal tolerance is
2
found in this genus, which underlines the fact that metal
responses may be strain-specific. As examples, P. lilacinum
comprises 23% of all fungi isolated from soil polluted by
mine drainage, and P. ochrochloron is commonly present in
industrial effluents.
In the plant kingdom, it seems clear that tolerance has
arisen independently in the full spectrum of families. It is
common to find species of Gramineae, Caryophyllaceae,
Lamiaceae and Fabaceae widely distributed along heavy-
metal contaminated ecosystems. The families Flacourtia-
ceae, Violaceae and Brassicaceae contain most of the
so-called hyperaccumulators, in which the genera Alyssum
(50 species) and Thlaspi (about 20 species) have been the
most widely characterized. Native vegetation from acid
soils of the humid tropics can grow in the presence of
concentrations of aluminium that are toxic to species not
associated with such habitats. Thus, organisms can evolve
mechanisms to cope with excess levels of heavy metals in
their environment.
In fact, hyperaccumulator plants have not only evolved
a mechanism to live in toxic concentrations of heavy
metals, but hyperaccumulation could also be functioning
as a strategy to prevent predation. As examples, the nickel-
rich leaves of Strephantus polygaloides prevent the devel-
opment of the herbivorous larvae of Pieris rapae and avoid
the growth of the phytopathogenic bacterium Xanthomo-
nas campestris. A repellent effect of the plant sap from
Sebertia acuminata (25% nickel dry wt) was observed on
the fruit fly Drosophila melanogaster (Sagner et al., 1998).
Mechanisms of Metal Uptake,
Exclusion, Transport and Storage
Uptake of some metal solutes from the soil environment
may occur through a carrier-mediated system, or as in the
case of cations, may be driven largely by the negative
potential across the plasma membrane, which is generated
in part by proton extrusion mediated by the membrane
enzyme H 1 ATPase.
Biological exudates, including microbial siderophores
and analogous compounds of plants termed phytosider-
ophores, are known to take part in the mobilization and
differential uptake of certain elements. In this way,
molybdenum and copper have been shown to form strong
complexes with this class of molecules, facilitating their
uptake.
Once in the cell, the organism must balance critically the
intracellular concentrations of these potentially toxic
metals. In some prokaryotes, animals and fungi, a class
of small cysteine-rich metal-binding proteins named
metallothioneins seems to be of primary importance in
metal compartmentalization and tolerance. These metal-
lothionein genes have been used to produce heavy-metal
tolerant transgenic plants. In plants, another class of

metal-binding ligands named phytochelatins has been
described. These proteins are rich in glutamine and
cysteine residues and may protect sensitive enzymes by
sequestering heavy metals, such as cadmium, lead and zinc
(Grill et al., 1987).
The majority of organisms that inhabit metalliferous
soils are known to exclude toxic metals. The production of
extracellular polysaccharides and the excretion of chelat-
ing substances, such as organic acids by microbial and
plant species, could participate in the immobilization of
such toxic elements. Another trait exploited by bacteria is
the accumulation of metals as cell-bound metal–phosphate
compounds. A number of plants, instead of excluding
heavy metals, have developed the unusual adaptation of
accumulating metals such as nickel, zinc or copper in their
above-ground biomass. These plants, termed hyperaccu-
mulators, have gained considerable attention owing to
their potential use in biorecovery of contaminated sites.
Little is known about the molecular, biochemical and
physiological processes that result in the hyperaccumula-
tor phenotype. However, low-molecular weight chelators,
such as certain amino acids and organic acids (e.g. citric),
have been shown to participate in the transport, compart-
mentalization and detoxification mechanisms.
In the genus Alyssum, in which the nickel concentration
can reach 3% of leaf dry biomass, the tolerant response
correlates with the increase in the levels of free histidine.
The supply of this amino acid to a nonaccumulating species
greatly increases both its nickel tolerance and transport to
the shoot (Kramer et al., 1996). However, the accumula-
tion of organic acids in roots and leaves of metal-tolerant
plants has also been described. Formation of anionic or
uncharged zinc–citrate complexes resulted in more zinc
passing through the excised stem of Pinus radiata and
copper in Papyrus stems. These complexes positively affect
transport in xylem by reducing adsorption to the vessel
walls and by decreasing the rate of lateral escape. A causal
relationship between organic acid accumulation and metal
tolerance has even been proposed. This is well established
for nickel-accumulating plants, which complex nickel with
malate, malonate and citrate. Apparently the organic acids
bind to heavy metals in the cytoplasm and this complex is
finally accumulated in the vacuole.
Another way in which organic acids may confer metal
tolerance has been found in some aluminium-tolerant
cultivars of snapbean, maize and wheat. In these plants, an
aluminium-resistant genotype correlates with the exuda-
tion of citrate and malate, and it has been shown that
organic acids prevent metal toxicity by chelating the
aluminium ions in the rhizosphere. This conclusion is
supported by the finding that the addition of organic acids
to toxic aluminium solutions abates aluminium toxicity in
the roots. Even more convincing is the fact that tobacco
and papaya transgenic plants, expressing a bacterial citrate
synthase gene and overproducing and overexuding citrate,
can grow in toxic concentration of aluminium by a
Heavy Metal Adaptation
presumptive metal exclusion mechanism (de la Fuente
et al., 1997).
Perspectives
Hyperaccumulator plants have been used as ‘indicators’
for metallic deposits for hundreds of years. Their
commercial importance for metal prospecting is indis-
putable. The shrub Hybanthus floribundus is used as an
indicator for nickel in Australia, while a wild variety of the
plant Impaticus balsamina, found on lead–zinc metal
dumps in India, is regarded as a local bioindicator for
these metals. In the lichen group, the species Cladonia
convoluta, which accumulates high copper concentrations
in its tissues, has become a tool for biogeochemical
prospecting and can be used to locate mining areas (Aery
and Tiagi, 1986).
The mechanism of metal hyperaccumulation is the
object of intensive investigations, especially with regard
to the ecological exploitation of such plants. Gradual
depletion of ecosystems encourages researchers not only to
investigate the natural response of organisms living on
contaminated sites, but also to suggest strategies for
environmental clean-up. The lack of affordable effective
approaches for heavy-metal remediation has created a
major need for the development of novel strategies. The
application of genetic engineering has been shown to be
highly successful in obtaining heavy-metal resistant
species. Transgenic plants tolerant to mercury, cadmium
and aluminium have recently been generated.
Although there has been some success with heavy-metal
resistant plants, the fact that we do not completely
understand the limiting factors in increasing uptake,
translocation and tolerance to toxic chemical elements
makes necessary the continuation of fundamental and
applied research. This should eventually make it possible
to overcome, at low monetary and environmental cost, the
problem of heavy metal contamination.
References
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