Environment International 26 (2001) 417 ± 423

www.elsevier.com/locate/envint

Relationships between chromium biomagnification ratio, accumulation

factor, and mycorrhizae in plants growing on tannery effluent-polluted soil
A.G. Khan*
School of Science, Food and Horticulture, College of Science, Technology and Environment, University of Western Sydney, Locked Bag 1797, Penrith South
DC, NSW 1797, Australia

Abstract

Heavy metal-contaminated land is increasingly becoming an important environmental, health, economic, and planning issue in Pakistan.
The unplanned disposal of industrial effluent from tannery, for example, has resulted in a many fold increase in chromium (Cr) in the land
near a tannery. This study was undertaken to compare the total and the DTPA-available Cr contents in the soil and the roots and leaves of tree
species growing on it with those on the nearby noncontaminated reference site at Kala Shah Kakoo, Panjab, Pakistan. A very reduced plant
cover on the tannery effluent-contaminated site was noted and there was a sharp boundary between the polluted and nonpolluted reference
sites, suggesting a strong selection pressure. Polluted soil contained considerable higher amounts of Cr as compared to the reference soil but
no correlation was found between Cr contents in the dried plant tissue and the total DTPA-extractable Cr. Roots of all the three tree species,
i.e. Dalbergia sissoo, Acacia arabica, and Populus euroamericana, growing on both the contaminated as well reference site possessed
arbuscular mycorrhizal fungal (AMF) infection in their roots and AMF propagules in the associated rhizospheres. D. sissoo and A. arabica
roots were also studded with nitrogen-fixing rhizobial root nodules, while those of P. euroamericana possessed AMF as well as
ectomycorrhizal infections. The dual infection would encourage mineral nutrition, including Cr. AMF community varied, i.e. trees growing
on the reference site were exposed to a wide variety of AMF such as Glomus, Scutellospora, and Acaulospora, whereas those on the
contaminated site contained only Gigaspora spp. in their mycorrhizospheres, suggesting a selection pressure. Typical Glomus infection
patterns in the roots of D. sissoo growing on the contaminated soil but absence of spores of Glomus spp. in the associated rhizospheres
indicate the potential error of using AMF spores to extrapolate the root infection. High Cr contents adversely affected the size, diversity, and
species richness of AMF as measured by Shannon ± Weiner index. The potential of mycorrhizae in protecting the host plant against the
harmful effect of heavy metals and in phytoremediation of the Cr-polluted soil is discussed. D 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Arbuscular mycorrhiza; Tannery effluent; Chromium; Phytoremediation; Rhizobial root nodules

1. Introduction
Soil ecosystems throughout the world have been con-
taminated with heavy metals by various human activities,
and movement of metals up the food chain has become a
human health hazard (Naidu et al., 1996). Heavy metal-
contaminated land is increasingly becoming an environ-
mental, health, economic, and planning issue in Pakistan
(Hussain et al., 1996). The rapid increase in population
together with the unplanned disposal of effluent from
tanneries and textile industries have increased the threat of
soil pollution in Pakistan. Studies by Pakistan EPA (EPA,
1990) have shown values for chromium (Cr) in the tannery
* Tel.: +61-2-4620-3237; fax: +61-2-4620-3025.
E-mail address: a.khan@uws.edu.au (A.G. Khan).
effluent, which are considerably higher than the safe limits
of the national standard. Basic Cr3 sulfate is used in the
tannery industry as the tanning agent because it forms
complexes with amino acids and proteins. Usually, uptake
of Cr into the leather is not complete and relatively large
amounts of Cr escape into the effluent. Furthermore, the
leather dust present in the effluent and other leather wastes
contain about 2% Cr, which reaches the environment as a
solid waste. All leather goods end up sooner or later in the
waste and Cr becomes more and more insoluble with time
due to formation of complexes with hydroxyl bridges.
Cr is an essential trace element, as it is required for
glucose metabolism and is found in food and feed in
concentrations between 0.05 and 2.4 mg/kg. Deficiency of
Cr in animals may cause diabetes, arteriosclerosis, growth
problems, and eye cataracts. Danger to its environmental
contamination depends on its oxidation state, i.e. hexavalent

0160-4120/01/$ ± see front matter D 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 1 6 0 - 4 1 2 0 ( 0 1 ) 0 0 0 2 2 - 8
418 A.G. Khan / Environment International 26 (2001) 417±423
Cr6 is more toxic than the common trivalent Cr3, which
precipitates at higher pH. Cr principally accumulates in the
surface layer (Mu et al., 1982) and exists in different forms
in soils. In a neutral alluvial soil contaminated with Cr3,
most of the Cr is bound by iron oxides and organic matter
and only a small proportion exists in the exchangeable form.
However, hexavalent (Cr6) form exists principally as anions
and its activities are controlled both by toxic minerals and
organic matter in these soils.
The damage to the environment caused by hazardous
tannery effluent is becoming an acute problem in Pakistan
and represents a technical challenge, as utilization of these
land for urban and/or agricultural developments requires a
safe and efficient decontamination process. For every 200
kg of leather, over 600 kg of solid waste is generated by a
tannery (Cabeza et al., 1998). Raju and Tandon (1999)
reported that 970 mg/g of Cr associated with the organic
phase in the sludge is formed as a result of basic tanning
activities. In 1995, the Supreme Court of India ordered the
closure of hundreds of tanneries in Tamil Nadu for failing to
treat their effluents (Kennedy, 1999). Various treatments
such as alkaline precipitation, reverse osmosis (Padilla and
Tavani, 1999), and activated sludge system (Ram et al.,
1999) have been proposed for treatment of Cr-contaminated
solid waste from a tannery. Such physiochemical techniques
are expensive and only suitable for small areas of high
commercial values. They are neither adaptable for specific
problems nor cost effective. Furthermore, the current phys-
iochemical methods render the land useless as a medium for
plant growth, as they remove all biological activities includ-
ing useful symbiotic microbes such as nitrogen-fixing
bacteria and mycorrhizal fungi as well as fauna in the
process of decontamination, thus decreasing their biodiver-
sity (Chaudhry et al., 1999).
New technologies are needed to address numerous con-
taminants, especially those that are neither volatile nor
mobile in soil solutions. One emerging technology, phytor-
emediation, is attracting increasing attention and is offering
a way to reclaim many of our urban sites lost to heavy metal
contamination. This technology employs plants with eco-
physiological adaptations to metalliferous soils to filter
(rhizofiltration) or absorb (phytoextraction/phytoaccumula-
tion) heavy metal(s) followed by their harvest (Chaudhry et
al., 1998). This technology is gaining popularity in reducing
metal load in the contaminated medium due to it being
environmentally friendly, less destructive to soil biota, and a
cheaper alternative. The major limitations to this powerful
technology include small biomass of plants adapted to grow
on nutrient poor-contaminated soils and disposal of the
heavy metal-enriched biomass.
Roots of most natural plants communities are occupied
by universal symbionts, i.e. arbuscular mycorrhizal (AM)
fungi, which enhance nutrient uptake including heavy
metals and promote the growth and survival of host plants
under nutrient-limiting conditions (Smith and Read, 1997).
AM fungi have been reported in plants growing on heavy
metal-contaminated soils (Khan et al., 2000), suggesting a
selective advantage for these plants as pioneering species
on such sites. A preliminary survey of the plants growing
on the tannery effluent-polluted soils at Kala Shah Kaku,
Pakistan showed that their roots were infected with sym-
biotic mycorrhizal fungi and studded with nitrogen-fixing
nodules (Khan and Bari 1997; Khan 1999a,b). Such plants
may be largely responsible for the successful colonization
of these habitats. Implications of such findings need to be
considered in the plant-based decontamination strategy,
i.e. phytoremediation.
The present study was undertaken to determine how the
diversity of AM fungi in the mycorrhizospheres of trees
growing on the above site is affected by the Cr-rich tannery
effluent. The principal aim was to assess the extent of
mycorrhization of roots of the tree species, the populations
of mycorrhizal fungal propagules in their associated rhizo-
spheres, and metal accumulation in their tissues. Some
mathematical indices such as Cr accumulation factor
[ACF = Cr in mature leaves (mg/g dw)/total Cr in soil (mg/
g dw)] and biomagnification ratio (BMR = Cr in mature
leaves and roots/available Cr in soil) values will also be
calculated to find out their relationships to mycorrhization.
2. Materials and methods
Samples of mycorrhizospheres soil, roots, and leaves of
three tree species, i.e. Populus euroamericana, Acacia
arabica, and Dalbergia sissoo, growing on the tannery
effluent-polluted soils around the tannery and on the nearby
noncontaminated reference site at Kala Shah Kaku, Panjab,
Pakistan were collected as per method of Crepin and
Johnson (1993). Soil pH and electrical conductivity (EC)
were measured. Mature leaves, roots, and rhizosphere soil
samples from five trees of the same species from each
polluted and reference site were collected, air-dried, and
mixed to form a composite sample. The soil, leaves, and
roots (except some root segments, which were retained
and fixed in FAA for determination of mycorrhizal infec-
tion) were dried for 2 days at 80°C and ashed in a muffle
furnace at 500°C for 6± 8 h. The ash was dissolved in a
mixture of 2 M HCl and 1 M HNO3 to determine total Cr
contents by atomic absorption spectrophotometry (Carter,
1993). The bioavailable fraction of the metals in the soil
samples was determined by the DTPA extraction method
(Lindsay and Norvell, 1978). The concentration of Cr was
expressed as mg/g of soil (Liang and Karamanous, 1993).
To illustrate plant species accumulatory ability, the ACF
and BMR were also calculated (Baker et al., 1994).
Washed roots were cleared and stained to determine
arbuscular mycorrhizal fungal (AMF) infection (Brundrett
et al., 1996). Presence or absence of functional nitrogen-
fixing nodules on the root pieces was also recorded.
Mycorrhizosphere soil samples were wet sieved and dec-
anted to recover AMF propagules (Tommercup, 1992). The
 A.G. Khan / Environment International 26 (2001) 417±423 419

density of spores in the soil and their species diversity were Table 2
Shannon ± Wiener index scores of AMF diversity (H) and AM fungal
determined. Different morphotypes of AMF spores recov-
species richness (R) as affected by Cr-rich tannery effluent contamination
ered were identified on the basis of spore size, colour, wall and different host plants
structure, and hyphal attachments to the species level
Shannon ± Weiner AMF
(Schenk and Perez, 1990; Morton and Benny, 1990). AMF diversity species
Several mathematical indices, i.e. AMF species richness Plant species Site index (H) richness (d)
(d), relative abundance of each species, and species diversity D. sissoo Reference 1.07 1.44
of AMF (H) in the mycorrhizospheres of the tree species D. sissoo Polluted 0.483 0.677
studied, were also determined (Krebs, 1985) to study the P. euroamericana Reference 1.246 1.394
effect of Cr on these parameters. P. euroamericana Polluted 0.451 0.926
A. arabica Reference 1.198 1.306
ANOVA was used to evaluate the effect of different host
A. arabica Polluted 0.655 0.782
trees and Cr contents on total spore number, species rich-
ness, and diversity of AM fungi. ANOVA was followed by
Duncan's test when appropriate. i.e. their cortices were occupied by relatively straight fungal
hyphae producing `H' branches characteristics of Glomus
infection, although no spores of Glomus spp. were found in
3. Results their mycorrhizospheres.
The root cortices of the trees growing on the nonpolluted
The EC and pH values of the tannery effluent-polluted reference site had mainly infection pattern of Glomus spp.,
soils were 26 dS m ÿ 1 and 8.5, respectively. A very reduced and Glomus spp. spores were recovered from their associ-
plant cover on the tannery effluent-contaminated site was ated rhizospheres. Some roots of P. euroamericana and D.
noted and there was a sharp boundary between the polluted sissoo from the reference site showed looping hyphae near
and nonpolluted reference sites. Roots of all the three tree entry points and no vesicles in their cortices, i.e. they were
species studied from both the contaminated and the refer- infected with Scutellospora spp. Examination of some roots
ence sites were found to be mycorrhizal. Ectomycorrhizal from the reference site revealed the presence of many
associations were observed in a few roots of P. euroamer- irregularly lobed intracellular oil-filled vesicles, a character-
icana. A 5± 7 mm thick mantle covered the fine rootlets. istic feature of Acaulospora spp. Very few arbuscules were
Root cortices of all the trees examined contained vesicles observed in the cortices of the roots examined. The roots of
and a few arbuscules, i.e. they possessed AM (Table 1). The D. sissoo and A. arabica growing on both the contaminated
infection ranges are 51± 71%, 34± 60%, and 70 ±80% in the and the reference sites had dual symbiotic associations, i.e.
roots of D. sissoo, P. euroamericana, and A. arabica, AMF infection as well as typical leguminous root nodules
respectively. The roots of the trees growing on the non- containing nitrogen-fixing Rhizobium bacteria (Table 1).
contaminated reference site were heavily mycorrhizal, i.e. As expected, a significantly higher number of AMF
>80%. The cortices of all the mycorrhizal roots of the trees propagules were recovered from the mycorrhizospheres
growing on the tannery effluent site contained thick walled samples collected from the reference site as compared to
and heavily stained fungal hyphae, intracellular oil-filled the contaminated site (Table 1). Cr contents in the soil also
vesicles, and intra- and intercellular-branched fungal hyphae exerted a differential effect on the AMF diversity index
with terminal vesicles, characteristics of infection by Giga- score (H) as measured by the Shannon± Wiener Index. A
spora spp. Some root pieces of D. sissoo growing in the lower level of diversity was observed in the Cr-polluted
contaminated soil also had typical Glomus spp. infection, soils than that in the rhizospheres of the trees growing on the

Table 1
DTPA extractable and total (g/g) Cr contents ( ‹ S.D.) of soil and the symbiotic associations AM, ectomycorrhizae (EM), and nitrogen-fixing root nodules (RN)
of the trees growing on the tannery effluent-polluted and reference soils
DTPA-extractable AM AMF spores
Tree species Cr Total Cr infection (%) (per 100 g soil) AM fungi Symbioses
D. sissoo (polluted) 200 ‹ 40 630 ‹ 88 51 ± 71 30 ‹ 5 Gigaspora spp. AM, RN
D. sissoo (reference) 80 ‹ 18 180 ‹ 26 >80 120 ‹ 22 Glomus, Acaulospora, AM, RN
and Scutellospora spp.
P. euroamericana 200 ‹ 40 630 ‹ 88 34 ± 60 12 ‹ 3 Gigaspora spp. AM, EM
(polluted)
P. euroamericana 80 ‹ 18 180 ‹ 26 >80 142 ‹ 34 Glomus, Acaulospora, AM, EM
(reference) and Scutellospora spp.
A. arabica (polluted) 200 ‹ 40 630 ‹ 88 70 ± 80 19 ‹ 5 Gigaspora spp. AM, RN
A. arabica (reference) 80 ‹ 18 180 ‹ 26 >80 198 ‹ 41 Glomus, Acaulospora, AM, RN
and Scutellospora spp.
Figures indicate the mean values of five samples significant at P > .05.
420 A.G. Khan / Environment International 26 (2001) 417±423
Table 3
Cr contents (mg/g dw) in root and shoot tissues of the trees growing on the
tannery effluent-contaminated soil and the calculated index values, Cr ACF
(ACF = Cr in mature leaves/total Cr in soil), and BMR (BMR = Cr in plant
tissue/available Cr in soil)
Cr in root Cr in leaf
tissue tissue ACF BMR
Tree species (mg/g dw) (mg/g dw) values values
D. sissoo 695 160 2.5 46.1
P. euroamericana 459 107 1.7 28.5
A. arabica 538 188 3.1 89.1
reference site (Table 2). The mycorrhizosphere soil samples
from the trees growing on the tannery effluent-polluted soils
were found to harbor AMF spores of Gigaspora strain only.
Many auxiliary vesicles, characteristics of Gigaspora, were
also present on the extraradical fungal hyphae attached to
the roots collected from the Cr-polluted site. This strain was
completely absent from the soil samples collected from the
mycorrhizospheres of the reference site, which contained
AMF spores of Glomus, Acaulospora, and Scutellospora
spp. (Table 1). The species richness (d) index scores of
AMF population was also affected by the tannery effluent,
i.e. d values decreased significantly in polluted soils, irre-
spective of the host trees (Table 2).
The DTPA extractable and total concentrations of Cr in
the soils from the polluted site were significantly higher than
the corresponding values in the reference soil ( P < .1; Table
1) and were in access of the soil quality guidelines of EPA,
Lahore (EPA, 1990). No significant correlation coefficient
was found between Cr contents in the dried plant tissue and
the total DTPA-extractable Cr in the soil. Calculated BMR
and ACF values were greater than 1. Cr ACF values ranged
from 1.7 to 3.1 and values for BMR ranged from 46.1 to
89.1 (Table 3). The derived accumulation factor based on
the total soil metal concentration values revealed the differ-
ing capacity of plant species for the accumulation of Cr in
the plant tissues (Table 3). The order of Cr accumulation
was D. sissoo >A. arabica >P. euroamericana. Mean values
of Cr accumulation (mg/g dw) in the leaves and the roots of
the three tree species studied from the contaminated site
showed that relatively higher amounts were retained by their
roots. A. arabica displayed higher levels of Cr as compared
to the other two tree species found on the tannery effluent-
polluted site. Based on the BMR and ACF ratios, A. arabica
was the most efficient accumulator of Cr, as it displayed the
highest values, i.e. BMR 89.1 and ACF 3.1.
4. Discussion
In the present study, major differences between the
contaminated and the reference sites in relation to the pH,
EC, and Cr contents (both total and available) were noted.
These differences accounted for the reduced plant cover on
the tannery effluent-contaminated site studied and suggest a
strong selection pressure. Heavy metal pollution may sup-
press or even kill sensitive parts of the plant and soil
microbial communities and lead to a shift in their functional
diversity and structure (FlieBbach et al., 1994; Chaudhry et
al., 1999).
The total Cr (630 mg/kg) in the tannery effluent-contami-
nated soils found in the present study is considerably less
than that reported by Raju and Tandon (1999) in the sludge
formed as a result of basic tanning activities (970 mg/kg) in
India. These authors also reported elevated concentrations
(1200 mg/kg) of Cr in plants growing on this sludge. On the
contrary, the present study did not reveal any significant
correlation between Cr content in the dried plant tissue and
the total DTPA-extractable Cr in the soil. In general, in spite
of high soil contents, only low concentrations of Cr were
found in the edible parts of the plants, e.g. spinach was
found to contain up to 23 mg/kg on dry weight basis (Kick
and Braun, 1977). However, some Australian trees such as
Leptospermum scoparium and Sutera fodina (Pimelea
suteri) were reported to accumulate much larger quantities
(2 ±4%) in the ash; Peterson and Girling, 1981). Wild (1974)
also reported much higher ash-weight Cr concentrations in
the leaves of Dicoma niccolefera and S. fodina from near a
chrome mine in Zimbabwe. The results of the present
studies, however, showed that relatively higher Cr amounts
were retained by the roots of the trees studied. The trans-
location of Cr from the root to the shoot is rather slow. It
tends to accumulate in root tissues and does not normally
correlate well with the extractable amounts in soil (Soon and
Abboud, 1993). Metal concentrations in aerial parts of
hyperaccumulators greatly exceed those of roots (Baker et
al., 2000). This may also be partly due to the method of
extraction used in this study. Shewry and Peterson (1976)
showed that acid ammonium oxalate was a better extractant
of soil Cr and it extracts a form of Cr, which is not readily
plant available and is different from that extracted by EDTA.
No speciation of Cr in the tannery effluent-contaminated
soil was determined in the present study. Hexavalent (Cr6),
however, is shown to easily cross the cell membranes
(Bianchi and Levis, 1984) where the phosphate ± sulfate
carrier also transports the Cr anions with a higher transport
index; Cr6 dominates more in the plant shoot (Smith et al.,
1989). Conversely, Cr3 does not utilize any specific mem-
brane carrier and its biological activity depends on both the
reduction and the subsequent storing in different compart-
ments and concentrates in the roots (Smith et al., 1989).
Furthermore, the question of biological availability of Cr
from soil for the plants is not clear. According to Herfeld
(1974), Cr from tannery wastes becomes more and more
insoluble with time by forming complex bridges. On the
contrary, Anderson (1977) suggested that after some period
of time, Cr may be fixed in the clay fraction of the soil. By
weathering, Cr can be set free again. A high Cr content of
plants growing on millions of years old serpentine soils
indicates that a certain amount of Cr is always available for
the plants (Sticher, 1978).
 A.G. Khan / Environment International 26 (2001) 417±423
The results of studies by the author demonstrate that
plants growing on the tannery effluent-contaminated soils
were mycorrhizal and that their mycorrhizospheres con-
tained AMF spores (Khan 1999a,b; Khan and Bari, 1997).
Mycorrhizae have been reported in plants growing on heavy
metal-contaminated sites (Khan et al., 2000), indicating that
these fungi have evolved heavy metal tolerance and that
they may play a role in the phytoremediation of the site. The
occurrence of only mycorrhizal trees on the Cr-contami-
nated site in the present study further suggests a selective
advantage for these trees as pioneering species on such sites
and that they may be largely responsible for the colonization
of such habitats.
The AMF community varied, i.e. trees growing on the
reference site are exposed to a wide variety of AM fungi
such as Glomus, Scutellospora, and Acaulospora, whereas
those on the contaminated site contained only Gigaspora
spp. in their rhizospheres. Sambandan et al. (1992) and
Raman and Sambandan (1998), on the contrary, reported
many AMF genera in tannery effluent-polluted soils of
Tamil Nadu, India. The AMF propagules were never dis-
appeared completely in the tannery effluent-polluted soils,
suggesting a certain adaptation of these indigenous AMF to
Cr. Various authors have reported isolation of spores of
AMF taxa from heavy metal-contaminated soils (Khan et
al., 2000) and changes in the diversity and abundance of
AMF populations (Koomen et al., 1990; Loth, 1996) due to
heavy metal contents. Other researchers found no correla-
tion between the concentration of heavy metals (Zn, Cu, Cd,
Ni, etc.) and AMF populations (Griffioen et al., 1994;
Weissenhorn and Leyval, 1994).
Typical infection pattern of Glomus spp. in the roots of
D. sissoo growing in the Cr-contaminated soils but
absence of spores of Glomus spp. in the associated rhizo-
spheres in the present study indicates the potential error of
using AMF spore surveys to extrapolate the root infection.
AMF may have identical morphological characteristics
inside and outside the roots; their function may differ
(Rosendahl et al., 1994). On the contrary, variations in
colour and size within a single isolate of Glomus clarum
were reported (Bentivenga et al., 1997). PCR fingerprint-
ing and ribosomal small subunit (SSU) sequences from
both AM root and AMF spore samples need to be
investigated to reveal genetic variations within AMF
species and genera (Clapp et al., 1999; Vandenkoornhuyse
and Leyval, 1998). However, mycorrhizal population
response to heavy metal toxicity as well as the biochem-
ical and molecular basis of tolerance is still poorly under-
stood (Turnau, 1993).
The complete absence of AMF spores of Gigaspora spp.
from the reference site and Glomus spp. in the polluted soils
in the present study indicates a selection pressure. Studies
by various workers have shown that mycorrhizal fungal
ecotypes from heavy metal-contaminated sites seem to be
more tolerant to heavy metals and have developed resistance
than reference strains from uncontaminated soils (Khan et
421
al., 2000). Galli et al. (1994) suggested that mycorrhizae can
play a crucial role in protecting plant roots from heavy
metals. More research is required to study the changes in the
genetic diversity of AM fungi.
The percentage of AM infection in the roots of trees
growing on the Cr-contaminated trees was found to be
reduced as compared to those growing on the reference site
in the present study. The extent of mycorrhization and the
quantity and quality of the AMF propagules in their rhizo-
spheres are clearly due to the Cr contents, which varied
significantly between the two sites. These results are in
agreement with those of Del Val et al. (1999) who have
shown that high concentrations of heavy metals adversely
affect the size, diversity, distribution, and activity of AMF
population in heavy metal-contaminated soils in a long-term
experiment. They also reported a reduced number of indig-
enous AMF propagules in heavy metal-contaminated soils
and a decrease in species richness and diversity as measured
by the Shannon± Weiner index.
The roots of all the trees of all the species examined
were heavily pigmented with phenolic and other secon-
dary metabolites and required bleaching before staining.
This may have reduced the subsequent staining of the
delicate hyphal branches of the arbuscules, explaining the
low frequencies of arbuscular infection observed. Con-
current occurrence of AM and ectomycorrhizae in the
roots of P. euroamericana observed in the present study
is in agreement with our previous observations (Chaudhry
et al., 1998) and indicates a lack of antagonism between
the two symbiotic fungi. Dual symbiosis of the AM with
root nodule inducing Rhizobia in Acacia and Dalbergia
spp. should further aid the adaptation, mineral nutrition,
and reduced root infection by pathogens of this plant
(Bethlenfalvay, 1992).
The values for the derived accumulation factor in the
present study revealed the differing capacity of the three tree
species studied. The uptake of Cr3 from soil depends upon
the plant species. Within a plant, the concentrations largely
differ between different parts of the plant (Sykes et al.,
1981). To further illustrate the accumulatory ability of the
trees growing on the Cr effluent-contaminated site in the
present study, a BMR was calculated and it was found that
A. arabica was the most efficient accumulator of Cr. Other
tree species might have evolved Cr exclusion strategy. The
response of different plant species to a particular heavy
metal varies markedly (Gabbrielli et al., 1990). On con-
taminated soils, growing side by side with metal accumu-
lator species, many species can be found, which are metal
tolerant and yet do not accumulate metal(s) (Brooks, 1987;
Reeves, 1992). The relationship between heavy metal accu-
mulation and tolerance may exist (Baker et al., 2000).
However, in the present study, no direct relationship
between Cr tolerance and accumulation was found. It is
important to note that in the present study, relatively higher
concentration of Cr was accumulated in the roots than in the
shoots, irrespective of the tree species studied. This indi-
422 A.G. Khan / Environment International 26 (2001) 417±423
cates that a different metal tolerance strategy, i.e. the
restriction of root-to-shoot transport of heavy metal, may
exist for metal tolerant nonaccumulators. Aggangan et al.
(1996) also found higher Cr and Ni values in the roots of
Eucalyptus urophylla seedlings inoculated with ectomycor-
rhizal fungus and grown in Cr-amended medium. These
results are in contrast with those of Nuruzzaman et al.
(1996) who reported a higher accumulation of Cr in the
above ground parts of Echinochloa crus-galli, a grass
growing in tannery effluent-contaminated soil in Bangla-
desh. Ectomycorrhizal fungi, such as Pisolithus, can
increase tolerance of trees to toxic metals (Jones and
Hutchinson, 1988) by accumulating metals in the extrama-
trical hyphae or by exclusion (Galli et al., 1994). Aggangan
et al. (1996) studied the effect of Cr on the growth of three
Pisolithus isolates and the formation of ectomycorrhizas on
E. urophylla seedlings in vitro and observed different
responses for the three isolates. These findings suggest that
screening of HM-tolerant mycorrhizal fungi could have
potential in phytoremediation.
Acknowledgments
I wish to thank Dr. Mohammad Ashraf Cheema, Director
General of Pakistan National Talent Pool, for providing me
an opportunity to undertake a TOKTEN mission in Pakistan
being financed by the United Nations (UNOP Grant no.
PAK/95/004) during 1997.
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