Environmental and Experimental Botany 71 (2011) 352–358

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Environmental and Experimental Botany

journal homepage: www.elsevier.com/locate/envexpbot

Characteristics of Cd uptake, translocation and accumulation in a novel

Cd-accumulating tree, star fruit (Averrhoa carambola L., Oxalidaceae)
J.T. Li, B. Liao, R. Zhu, Z.Y. Dai, C.Y. Lan, W.S. Shu ∗
School of Life Sciences and State Key Laboratory of Biocontrol, Sun Yat-Sen University, No. 135, Xing-Gang-Xi Road, Guangzhou 510275, PR China

a r t i c l e i n f o a b s t r a c t

Article history: Cadmium (Cd) accumulation by terrestrial higher plants is an intriguing phenomenon that may be
Received 20 December 2009 exploited for phytoextraction of Cd-contaminated soils. Characterizing the physiological processes
Received in revised form responsible for elevated concentrations of Cd in shoots is a first step towards a comprehensive under-
23 December 2010
standing of the mechanisms underlying Cd accumulation in plants and may eventually improve the
Accepted 2 February 2011
efficiency of phytoextraction. Woody species that can accumulate Cd have been recently recommended
as good candidates for phytoextraction of Cd-contaminated soils. However, little is known about the
Keywords:
mechanisms of Cd accumulation by woody species. In an attempt to understand the physiological pro-
Averrhoa carambola
Cd hyperaccumulation
cesses contributing to Cd accumulation in woody species, Cd uptake and translocation by a novel tropical
Translocation Cd-accumulating tree, star fruit (Averrhoa carambola) were characterized and compared with those of
Uptake a non-Cd-accumulating tree (Clausena lansium). Our results showed that A. carambola had higher Cd
Zn pathway uptake and root-to-shoot translocation efficiencies than C. lansium, which might account for its greater
Cd-accumulating capacity. Furthermore, Cd accumulation by A. carambola was not significantly affected
by zinc (Zn), whereas Zn accumulation was greatly lowered by Cd. This phenomenon could not be fully
explained by a simple competition between Cd2+ and Zn2+ , implying the existence of a transport sys-
tem with a preference for Cd over Zn. Collectively, our results indicate that A. carambola has noteworthy
physiological traits associated with accumulation of Cd to high levels.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction
Cadmium (Cd) is regarded as one of the most toxic environ-
mental pollutants and is readily absorbed by certain crops from
soils (Wagner, 1993). Elevated concentrations of Cd in agricul-
tural soils have posed a significant threat to safe food production
and have therefore become a worldwide concern (Wagner, 1993;
McLaughlin et al., 1999; Järup and Åkesson, 2009). The last two
decades have witnessed an increasing need for the remediation of
Cd-contaminated soils (Maejima et al., 2007; Leštan et al., 2008).
Phytoextraction, i.e. the use of plants to remove pollutants
(including Cd) from contaminated soils, has been proposed as a
promising technology that is both low cost and environmentally
friendly (Baker et al., 2000). A general approach for phytoextraction
of Cd-contaminated soils is to make use of Cd-hyperaccumulating
plants (Baker et al., 2000; McGrath and Zhao, 2003). However,
to date, only a few herbaceous species have been identified as
Cd-hyperaccumulators, including Thlaspi caerulescens, Arabidopsis
halleri, Viola baoshanensis, Sedum alfredii, Sedum plumbizincicola and
∗ Corresponding author. Tel.: +86 20 39332933; fax: +86 20 39332944.
E-mail addresses: shuws@mail.sysu.edu.cn, zsushuwensheng@163.com
(W.S. Shu).
Thlaspi praecox (Baker et al., 1994; Bert et al., 2002; Liu et al., 2004;
Yang et al., 2004; Jiang et al., 2010; Vogel-Mikuš et al., 2010). Fur-
thermore, in most cases, hyperaccumulators tend to grow slowly
and produce a relatively low biomass (Krämer, 2010), which greatly
limits the speed of Cd removal (McGrath et al., 2006; Wu et al., 2007;
Zhuang et al., 2007).
In recent years, interest has grown in the potential use of
woody species for metal phytoextraction (Pulford and Watson,
2003; Dickinson and Pulford, 2005; Mertens et al., 2006; Komárek
et al., 2008; Jensen et al., 2009). Woody species generally have
high biomass and extensive root systems, which make them poten-
tially suitable for phytoextraction (Pulford and Watson, 2003). In
addition, a dense tree canopy can also prevent the spread of con-
taminated soil by wind erosion. There are a very few naturally
occurring Cd-accumulating trees identified so far that may be effi-
cient at removing Cd from contaminated soils, such as Salix caprea,
Salix fragilis and Salix striandra (Mertens et al., 2006; Wieshammer
et al., 2007). However, for most of the other woody species tested,
their capacities to extract Cd from contaminated soils are rela-
tively low, which makes their large-scale applications currently
non-feasible (van Nevel et al., 2007; Jensen et al., 2009; Wang and
Jia, 2010).
One possible strategy to improve the efficiency of Cd phytoex-
traction would be the transfer of appropriate genes involved in Cd

0098-8472/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.envexpbot.2011.02.001
 J.T. Li et al. / Environmental and Experimental Botany 71 (2011) 352–358
accumulation to fast-growing and high-biomass species (Cherian
and Oliveira, 2005; Doty, 2008). This avenue, however, requires
a greater understanding of mechanisms underlying Cd accumu-
lation in plants (Verbruggen et al., 2009). Root uptake of Cd and
root-to-shoot Cd translocation are the two key physiological pro-
cesses related to Cd accumulation in plants (Clemens, 2006). A
better understanding of these processes is therefore essential for
a comprehensive understanding of the mechanisms underlying
Cd accumulation in plants (Clemens, 2006). There is increasing
evidence that enhanced root uptake and/or root-to-shoot translo-
cation of Cd are responsible for the high accumulation of Cd in
T. caerulescens, A. halleri and S. alfredii (Lombi et al., 2001; Zhao
et al., 2002, 2006; Lu et al., 2008). For example, Zhao et al. (2002)
found that symplastic root uptake of Cd is 4.5 times higher in
the high Cd-accumulating T. caerulescens ecotype (Ganges) than in
the low Cd-accumulating ecotype (Prayon), while Lu et al. (2008)
reported that the rate of root-to-shoot translocation of Cd in a
hyperaccumulating S. alfredii ecotype is >10-fold higher than in
a non-hyperaccumulating ecotype. On the other hand, no high-
affinity Cd transporter has been identified, although evidence has
shown that radial symplastic passage and rapid xylem loading are
required for efficient Cd uptake and translocation in the plants
(Verbruggen et al., 2009). Alternatively, Cd uptake and translo-
cation in Cd-accumulating plants may at least partly be achieved
through the Zn pathway (Zhao et al., 2002, 2006; Wong and Cobbett,
2008; Verbruggen et al., 2009). The significant progress mentioned
above was made in Cd-accumulating herbaceous species, whereas
little is known about the physiology of Cd accumulation in woody
species.
Averrhoa carambola L., Oxalidaceae (star fruit) is a novel tropical
Cd-accumulating tree, which has potential for Cd phytoextraction
(Li et al., 2009). This tree can accumulate considerable amounts
of Cd in its shoots (approximately 500 mg kg−1 , on a dry weight
basis, DW) without significant biomass reduction, when grow-
ing in solutions containing 5 mg Cd L−1 for 3 months (Li et al.,
2010). In the present study, the characteristics of root uptake and
root-to-shoot translocation of Cd in A. carambola were investi-
gated and compared with those of a non-Cd-accumulating tree
[Clausena lansium (Lour.) Skeels, Rutaceae], to unravel the physi-
ological processes contributing to the Cd-accumulating capacity of
this woody species. C. lansium was selected to be compared with
A. carambola; because this tree is a non-Cd-accumulating species
(Li et al., 2006), while most close relatives of A. carambola are
herbaceous (Huang et al., 1998). Interactions between transport
of Cd and Zn in A. carambola were characterized to determine
whether or not the root uptake and root-to-shoot transloca-
tion of Cd in this species were mediated via transporters for
Zn.
As root uptake of Cd is the first step of Cd accumulation in plants
(Clemens, 2006), the characteristics of root uptake have frequently
been investigated to explain the differences in Cd-accumulating
capacity between species, ecotypes and cultivars (Piñeros et al.,
1998; Lombi et al., 2001; Zhao et al., 2002; Lu et al., 2008; Redjala
et al., 2009). The previous studies have shown that root uptake of
Cd can be partitioned into an apoplastic and a symplastic compo-
nent, which represent Cd binding to cell walls and trans-plasma
membrane transport of Cd, respectively. It is therefore impor-
tant to distinguish the symplastic Cd fraction from the apoplastic
one in the characterization of Cd influx into roots. One widely
used approach for estimating Cd binding in root apoplast is to
determine the apparent uptake of Cd at 2 ◦ C, assuming that sym-
plastic root uptake of Cd is negligible at this low temperature
(Lombi et al., 2001; Zhao et al., 2002; Lu et al., 2008; Redjala
et al., 2009). In the present study, this approach was therefore
applied to characterize apoplastic Cd binding in roots of the two
species.
353
2. Materials and methods
2.1. Plant material and culture conditions
Uniform 3-month-old seedlings of A. carambola (cv. Malaysia)
and C. lansium (cv. Ji-Xin) were provided by the Guangzhou Insti-
tute of Fruit Sciences (Guangzhou, China). For hydroponic cultures,
seedlings were transferred to 5.0 L vessels containing basal nutrient
solution and placed in a glasshouse (25 ± 1 ◦ C). The composition of
the basal nutrient solution was as follows: 2.5 mM KNO3 , 2.5 mM
Ca(NO3 )2 , 1 mM MgSO4 , 0.5 mM KH2 PO4 , 100 ␮M H3 BO3 , 40 ␮M
MnCl2 , 2 ␮M CuSO4 , 1 ␮M ZnCl2 , 0.4 ␮M MoO4 Na2 , and 100 ␮M
NaFe-EDTA. The pH of the solution was maintained at 6.0 ± 0.1
using HCl or NaOH, as required. The nutrient solution was renewed
weekly. After 2 weeks of preculture, healthy seedlings with new
roots were used for further experiments.
2.2. Cd-uptake experiments
Cadmium uptake by roots of A. carambola and C. lansium
was investigated using a depletion technique similar to that
previously described by Zhao et al. (2002). In detail, seedlings
were pretreated with a solution containing 2 mM MES (2-
morpholinoethanesulphonic acid, pH 6.0) and 0.5 mM CaCl2 for
24 h before the uptake experiment. The uptake experiment was
initiated at 2 h after the beginning of the light period and per-
formed at 25 ± 1 ◦ C. The seedlings were transferred to 125 mL
vessels (one seedling per vessel) filled with 100 mL of uptake solu-
tion containing 2 mM MES (pH 6.0), 0.5 mM CaCl2 and 10 ␮M CdCl2 .
For each species, 4 replications were set up and each replication
comprised 1 seedling. In order to monitor the depletion of Cd in
uptake solutions, at 0, 0.5, 1, 2, 3, 4 h after the initiation of uptake
experiment, 1.0 mL solution was taken from each vessel for Cd
determination. The removed solutions were compensated by addi-
tion of deionized water. At the end of the uptake experiment,
roots of the seedlings were collected and weighed. Concentra-
tions of Cd in the roots were calculated from the depletion of
Cd in the uptake solutions and expressed on a fresh weight basis
(FW). For example, concentration of Cd in the root exposed to Cd
for 1 h (C1 h-root ) could be calculated according to this equation:
C1 h-root = [(C0 h-solution − C1 h-solution ) × the total volume of uptake
solution]/root weight, where C0 h-solution stands for the initial Cd
concentration in uptake solution and C1 h-solution is the Cd concen-
tration in uptake solution sampled at 1 h after the initiation of
uptake experiment. The root uptake of Cd at 25 ◦ C represents the
total amount of Cd taken by the roots, which comprises apoplastic
and symplastic components (Zhao et al., 2002).
To estimate the apoplastic Cd binding in roots of the two woody
species, the uptake experiment described above was also carried
out at 2 ◦ C (Zhao et al., 2002), in which the seedlings were trans-
ferred to an ice-cold (2 ◦ C) pretreatment solution for 30 min and
then kept in vessels that contained 100 mL uptake solution. During
the 4-h uptake experiment, the vessels were placed in an ice bath
and shaded from light. The other experimental procedures were
the same as those used previously at 25 ± 1 ◦ C.
2.3. Cd-translocation experiment
To investigate Cd translocation from roots to shoots, time-
course experiments involving Cd accumulation in the shoots were
conducted according to Xing et al. (2008). Seedlings of the two
species were transferred to 2.0 L vessels (four seedlings per vessel)
and pretreated with the basal nutrient solutions spiked with 10 ␮M
CdCl2 for 24 h. Four replications were set up for each species and
each replication comprised 4 seedlings. After pretreatment with
Cd for 24 h, the roots of the seedlings were immersed into the basal
354 J.T. Li et al. / Environmental and Experimental Botany 71 (2011) 352–358
nutrient solutions for 20 min to remove Cd from the root surfaces.
Following this procedure, 4 seedlings (one seedling from each repli-
cation) of each species were harvested and the others were grown
in the basal nutrient solutions without Cd supplement. For each
species, 4 seedlings were harvested after culture in the Cd-free
nutrient solutions for 1, 5 and 9 days, respectively. After harvest,
all plants were rinsed with deionized water; twigs (twigs + stems)
and leaves were oven-dried at 70 ◦ C to a constant weight and then
their weights were recorded.
2.4. Interactions between transport of Cd and Zn
To determine whether A. carambola accumulates Cd through the
Zn pathway, the interactions between Cd and Zn transport were
characterized using the method described by Zhao et al. (2006).
To test the effects of Cd on Zn transport in A. carambola, seedlings
were exposed to 2.0 L basal nutrient solutions containing 0 (con-
trol), 1, 5, 15 or 50 ␮M CdCl2 and a constant 5 ␮M Zn(NO3 )2 .
Likewise, seedlings of A. carambola were grown in 2.0 L basal nutri-
ent solutions containing 1 (control), 5, 50 or 500 ␮M Zn(NO3 )2
and a constant 5 ␮M CdCl2 , to investigate the effects of Zn on Cd
accumulation. Each treatment was replicated 4 times and each
replication comprised 4 seedlings in a vessel. All nutrient solu-
tions were renewed weekly. The vessels were placed randomly
in a glasshouse (25 ± 1 ◦ C) and repositioned once a week. After 8
weeks of growth, all plants were harvested and divided into roots,
twigs (twigs + stems) and leaves. The 4 seedlings per vessel were
combined into one sample. To remove the Cd adsorbed onto root
surfaces, roots of each seedling were immersed into 20 mM Na2 -
EDTA (disodium ethylenediaminetetraacetate) solutions for 0.5 h.
The plant materials were then treated as described above. In addi-
tion, the dry biomasses of different tissues were weighed.
2.5. Chemical analyses
For heavy metal analyses, dried plant material was ground and
digested with a mixture of HNO3 /HClO4 (5:1, v/v). Concentrations
of the metals in digests or uptake solutions were measured using a
graphite furnace atomic absorption spectrophotometer (Hitachi Z-
5300; Tokyo, Japan). To investigate, preliminarily, the interactions
between transport of Cd, calcium (Ca) and iron (Fe) in A. carambola,
Ca and Fe concentrations in different plant tissues from experi-
ments regarding the interactions between transport of Cd and Zn
were also determined.
2.6. Statistical analysis
Statistical analysis was performed using SPSS version 16.0 for
windows (SPSS Inc., USA). Student’s t-test was used to assess sig-
nificant differences in Cd uptake and translocation between A.
carambola and C. lansium. In the experiments concerning interac-
tions between transport of Cd and Zn in A. carambola, we measured
substantial variations in biomass production and tissue heavy
metal concentrations among Cd or Zn treatments by a one-way
ANOVA followed by Fisher’s least significant difference (LSD) test.
3. Results
3.1. Time-dependent Cd uptake by roots
The symplastic root uptake of Cd in A. carambola and C. lansium
was calculated by subtracting apoplastic Cd binding in root (root
uptake of Cd at 2 ◦ C) from the total Cd uptake in roots (root uptake
of Cd at 25 ◦ C). During the 4-h uptake experiment, the symplastic
root uptake of Cd in the two species increased gradually with time
(Fig. 1). However, the symplastic root uptake of Cd in A. carambola
7
SU-Cd in A. carambola
SU-Cd in C. lansium
Cd concentrations in roots (μmol g -1 , FW)
6 AB-Cd in A. carambola *
AB-Cd in C. lansium
*
5
*
4
3
2
*
1
0
0 1 2 3 4
Time (h)
Fig. 1. Time-dependent symplastic root uptake of Cd (SU-Cd) and apoplastic Cd
binding in root (AB-Cd) of A. carambola and C. lansium treated with 10 ␮M CdCl2 for
4 h. Data are presented as means ± S.D. (n = 4). Asterisks indicate that the root SU-Cd
in A. carambola is significantly (P < 0.05, t-test) higher than that of C. lansium at the
same time.
was consistently higher than that of C. lansium; significant (P < 0.05)
differences between the two species were found at all times after
1 h of uptake (Fig. 1). At the end of the uptake experiment, the sym-
plastic root uptake of Cd was 2.1 times higher in A. carambola than
in C. lansium. In contrast, there were no significant (P > 0.05) dif-
ferences between the two species in apoplastic Cd binding in roots
(Fig. 1).
3.2. Time-dependent Cd accumulation in shoots
After pretreatment with Cd solutions for 24 h (day 0 in Fig. 2),
the Cd concentrations in twigs and leaves of A. carambola were 2.96
and 3.16 mg kg−1 (DW), being 3.5 and 3.3 times higher than those in
C. lansium, respectively (Fig. 2). Afterwards, the Cd concentrations
in twigs and leaves of A. carambola increased rapidly with time
over the following 9 days. However, those in C. lansium increased
very slowly (Fig. 2). Consequently, on day 9 after culture in Cd-free
solutions, the Cd concentrations in twigs and leaves of A. carambola
were 4.5 and 9.8 times higher than those in C. lansium, respectively
(Fig. 2).
3.3. The effects of Cd on Zn accumulation in A. carambola
Increasing Cd concentrations in solutions with a constant con-
centration of Zn (5 ␮M) did not significantly decrease biomasses
of A. carambola; instead, a significant increase in root biomass at
15 ␮M Cd treatment was recorded (Fig. 3a). The Cd concentrations
in different tissues of A. carambola increased significantly (P < 0.05)
with increasing solution Cd, while the highest tissue Cd concen-
tration (469 mg kg−1 , DW) was recorded in the leaves at 50 ␮M Cd
(Fig. 4a). By contrast, the Zn concentrations in different tissues of
A. carambola were significantly decreased (P < 0.05) by increasing
Cd concentrations in solutions (Fig. 4b). The Zn concentrations in
roots, twigs and leaves were 1.9, 3.1 and 3.7 times lower at 50 ␮M
Cd than at 0 ␮M Cd, respectively.
 J.T. Li et al. / Environmental and Experimental Botany 71 (2011) 352–358 355

8 1.0 (a) The effects of Cd treatments

(a) Twigs
Cd concentration (mg kg , DW) ***
A. carambola 0 μM Cd 1 μM Cd 5 μM Cd 15 μM Cd 50 μM Cd
-1

Tissue biomass (g plant , DW)

6 0.8
C. lansium
a a
a a a

-1
***
4 ** 0.6 b b a
b b a a a a a
**

0.4
2

0.2
0
12
(b) Leaves
Cd concentration (mg kg , DW)

***
10 0.0
1.0
(b) The effects of Zn treatments
-1

Tissue biomass (g plant , DW)

1 μM Zn 5 μM Zn 50 μM Zn 500 μM Zn
0.8
6
** a

-1
a a
* ** a
4 0.6 a a a a
a a a
b
2
0.4
0
0 2 4 6 8 10
0.2
Time (day)

Fig. 2. Time-dependent Cd accumulation in A. carambola and C. lansium that were

pretreated with 10 ␮M CdCl2 for 24 h and then grown in basal nutrient solutions
without Cd supplement. Data are presented as means ± S.D. (n = 4). Asterisks indi-
cate that significant differences between the two species: *P < 0.05; **P < 0.01 and
***P < 0.001 (t-test).
3.4. The effects of Zn on Cd accumulation in A. carambola
Tissue biomass of A. carambola was not significantly (P > 0.05)
reduced by increasing Zn in solutions with a constant concentra-
tion of Cd (5 ␮M), with the only exception being the root biomass
obtained in the 500 ␮M Zn treatment (Fig. 3b). Increasing solution
Zn resulted in significant (P < 0.05) increases in Zn concentrations in
tissues of A. carambola, while the maximum tissue Zn concentration
(2121 mg kg−1 , DW) was obtained in leaves at 500 ␮M Zn treat-
ment (Fig. 5a). On the other hand, 5 and 50 ␮M Zn slightly (P > 0.05)
increased the Cd concentrations in different tissues of A. carambola,
whereas the 500 ␮M Zn treatment significantly (P < 0.05) decreased
the tissue Cd concentrations (Fig. 5b).
3.5. The effects of Cd on Ca and Fe accumulation in A. carambola
Concentrations of Ca and Fe in different tissues of A. carambola
were in the normal ranges for plants (Fig. 6). On the other hand,
they were not significantly (P > 0.05) influenced by the increasing
Cd levels in nutrient solutions (Fig. 6, details on results of LSD tests
not shown).
4. Discussion
In the present study, based on the results of the uptake exper-
iment at 2 ◦ C, there were no significant differences between the
Cd-accumulator A. carambola and the non-accumulator C. lansium
in apoplastic Cd binding in roots (Fig. 1). Similarly, no significant
difference was found between the two contrasting T. caerulescens
ecotypes Ganges and Prayon in Cd binding in root apoplast (Zhao
et al., 2002). However, an inconsistent result was recently reported
by Lu et al. (2008) who found that the apoplastic Cd binding in root
0.0
Roots Twigs Leaves
Fig. 3. The effects of Cd (a) and Zn (b) treatments on tissue biomass of A. carambola
grown in Cd or Zn solutions for 8 weeks. Data are presented as means ± S.D. (n = 4).
Bars marked by the same letters do not differ significantly (LSD, P < 0.05).
is greater in the non-Cd-hyperaccumulating S. alfredii ecotype than
in the Cd-hyperaccumulating one. These results suggest that there
is no consistent link between apoplastic Cd binding in roots and the
Cd-accumulating capacity of plants. In contrast, it has always been
found that Cd-hyperaccumulating plants take up more Cd sym-
plastically into roots than non-Cd-hyperaccumulating ones (Lombi
et al., 2001; Zhao et al., 2002; Lu et al., 2008). In line with the pre-
vious results, the present study showed that the time-dependent
symplastic root uptake of Cd was significantly greater in the Cd-
accumulator A. carambola than in the non-accumulator C. lansium
(Fig. 1). This result implies that the greater Cd-accumulating capac-
ity of A. carambola compared to C. lansium might, at least partly, be
attributed to the greater symplastic root uptake of Cd in A. caram-
bola.
Cadmium accumulation in shoots is a direct result of root-to-
shoot translocation of Cd, which is an important physiological
process subsequent to Cd uptake into the root symplasm (Clemens,
2006). It is well known that Cd-hyperaccumulators are charac-
terized by high Cd translocation factors (the shoot-to-root Cd
quotients > 1). However, there are contrasting opinions about the
extent to which Cd hyperaccumulation in plants can be explained
by their more efficient root-to-shoot translocation of Cd. Lombi
et al. (2001) found that the concentration of Cd in xylem sap col-
lected from T. caerulescens Ganges ecotype is 5 times higher than
that of Prayon. These authors, however, believed that this differ-
ence was largely ascribed to the 5 times difference in root uptake
of Cd between the two ecotypes. Therefore, they concluded that
the divergence between the two T. caerulescens ecotypes in Cd-
accumulating capacity is largely explained by their difference in
root uptake of Cd, rather than in translocation of Cd from root
to shoot (Lombi et al., 2001). In contrast, Lu et al. (2008) found
356
1000
Cd concentration (mg kg , DW) (a) Cd
-1
100
10
1
1000 (b) Zn
Zn concentration (mg kg , DW)
-1
100
0 1 5 15
Solution Cd (μM)
Fig. 4. The effects of solution Cd on Cd (a) and Zn (b) concentrations in different
tissues of A. carambola grown in solutions with 0, 1, 5, 15 or 50 ␮M Cd, and a constant
5 ␮M Zn for 8 weeks. Data are presented as means ± S.D. (n = 4).
that there is 46 times higher concentration of Cd in shoots of
a Cd-hyperaccumulating S. alfredii ecotype than in the non-Cd-
hyperaccumulating one, which cannot be explained by the twice
higher symplastic root uptake of Cd in the Cd-hyperaccumulating
S. alfredii ecotype. On this basis, it was concluded that Cd hyper-
accumulation in the Cd-hyperaccumulating S. alfredii ecotype was
largely attributed to greater root-to-shoot translocation of Cd,
rather than enhanced symplastic root uptake of Cd (Lu et al., 2008).
Interestingly, our results suggest that the greater Cd-accumulating
capacity of A. carambola compared with C. lansium can be ascribed
not only to the enhanced symplastic root uptake of Cd, but
also to the greater root-to-shoot translocation of Cd. In the Cd-
translocation experiment of our study, the Cd concentrations in
twigs and leaves of A. carambola on day 0 were 3.5 and 3.3 times
higher than those of C. lansium, respectively (Fig. 2). It is unlikely
that these about 3 times differences between the two species in
shoot Cd concentrations would change greatly in the following 9
days without further supply of Cd in solutions, if there was no dif-
ference between the two species in root-to-shoot translocation of
Cd. However, at the end of the Cd-translocation experiment (day
9, Fig. 2), there were 4.5 and 9.8 times higher concentrations of
Cd in twigs and leaves of A. carambola than in those of C. lansium
which could not be fully explained by the twice higher symplas-
tic root uptake of Cd in A. carambola (Fig. 1). These results show
that the root-to-shoot translocation of Cd was more pronounced in
A. carambola than in C. lansium, which may be due to more xylem
loading of Cd (Clemens, 2006).
No high-affinity Cd transporter has been identified, although a
growing body of physiological evidence suggests that more efficient
root uptake and/or root-to-shoot translocation of Cd have evolved
in Cd-accumulating plants (Verbruggen et al., 2009). Alternatively,
J.T. Li et al. / Environmental and Experimental Botany 71 (2011) 352–358
(a) Zn
Roots
Zn concentration (mg kg , DW)
Twigs
Leaves
1000
-1
Roots
100
Twigs
Leaves
10
1000
Roots (b) Cd
Twigs Roots
Cd concentration (mg kg , DW)
Leaves Twigs
Leaves
-1
100
50
1 5 50 500
Solution Zn (μM)
Fig. 5. The effects of solution Zn on Zn (a) and Cd (b) concentrations in different
tissues of A. carambola grown in solutions with 1, 5, 50 or 500 ␮M Zn, and a constant
5 ␮M Cd for 8 weeks. Data are presented as means ± S.D. (n = 4).
as a non-essential but highly toxic element for plant growth, Cd has
always been demonstrated to be transported in plants by trans-
porters for essential elements, especially for Zn (Clemens, 2006).
Zhao et al. (2006) found that the Cd hyperaccumulation in A. hal-
leri is at least partly mediated by the Zn pathway. However, it
was recently reported that Cd uptake by a Cd-hyperaccumulating
S. alfredii ecotype is likely regulated by Ca transporters/channels
rather than Zn transporters (Lu et al., 2008). In the present study,
the possibility that Cd accumulation in A. carambola was also
related to the Zn pathway was tested, in order to shed light on
the role of Zn transporters in Cd accumulation in woody species.
Our results showed that increasing Cd concentrations in solutions
significantly decreased the Zn concentrations in different tissues,
while they greatly increased Cd concentrations in tissues of A.
carambola (Fig. 4). At first glance, this phenomenon was not sur-
prising, because, the decreased tissue Zn concentrations might
largely be explained by the well-documented competitive inter-
actions between Cd and Zn ions for entry into the active sites of
Zn transporters (Clemens, 2006). The increased Cd concentrations
in the tissues could mainly be related to the increases in the con-
centrations of free Cd ions in corresponding solutions (Zhao et al.,
2006). However, upon examination of Zn effects on Cd transport in
A. carambola, it was interesting to find that the interactions between
transport of Cd and Zn in this species could not be fully inter-
preted by simple competition between the two ions. In contrast to
the expectation, Cd concentrations in tissues of A. carambola were
slightly increased by increasing concentrations of Zn in solutions
(Fig. 5), with the exception of decreases recorded at the highest Zn
treatment (500 ␮M Zn) which might be attributed to toxicity of Zn
in this species, considering that the root biomass of A. carambola at
 J.T. Li et al. / Environmental and Experimental Botany 71 (2011) 352–358
28000 (a) Ca
Roots
Ca concentration (mg kg , DW)
24000 Twigs
Leaves
-1
20000
16000
12000
8000
4000
1000 (b) Fe
Fe concentration (mg kg , DW)
-1
100
Roots
Twigs
Leaves
10
0 1 5 15
Solution Cd (μM)
Fig. 6. The effects of solution Cd on Ca (a) and Fe (b) concentrations in different
tissues of A. carambola grown in solutions with 0, 1, 5, 15 or 50 ␮M Cd, and a constant
5 ␮M Zn for 8 weeks. Data are presented as means ± S.D. (n = 4).
the highest Zn treatment was significantly lowered (Fig. 3b). This
result was distinct from those of previous studies showing that Cd
concentrations in tissues of the Cd-hyperaccumulator A. halleri and
non-accumulating crops (such as Triticum aestivum and Triticum
turgidum) are significantly lowered by increasing solution Zn (Hart
et al., 2002; Zhao et al., 2006). There are two possible explanations
for this interesting finding of our study, although the elaborate
underlying mechanisms remain to be demonstrated. The first is
that upon exposure to increasing Zn concentrations in solutions,
the expression and/or activity of Zn transporters in A. carambola
is enhanced which is sufficient for transport of both Zn and Cd
ions from the solution to tissues. However, most available evidence
argues against such a possibility. As reviewed by Verbruggen et al.
(2009), most identified transporters involved in Zn/Cd uptake and
translocation (such as ZNT1 and AtHMA4) are generally expressed
independently of the Zn supply in hyperaccumulators, and induced
by Zn deficiency in non-accumulators. An alternative explanation
is the existence of a transport system with a preference for Cd over
Zn. As seen in Fig. 5b, Cd concentrations in tissues were not sig-
nificantly lowered, even when the molar ratio of solution Zn to Cd
reached 10, suggesting that Cd was bound better than or at least as
good as Zn to a transporter. Similarly, a putative Cd-preferred trans-
port system was assumed to function in Cd hyperaccumulation in
the Ganges population of T. caerulescens (Lombi et al., 2001; Zhao
et al., 2002). In addition, it was found that the increasing solution
Cd did not significantly (P > 0.05) alter the Ca and Fe concentrations
in tissues of A. carambola (Fig. 6), indicating that the transport sys-
tems for Ca and Fe were not involved in Cd accumulation in this
species (Clemens, 2006; Dong et al., 2006).
357
5. Conclusions
This study showed that the root uptake and root-to-shoot
translocation of Cd in A. carambola were more efficient than those
in C. lansium, which possibly contributed to the Cd-accumulating
capacity of A. carambola. Furthermore, it was found that Cd trans-
port in A. carambola was not significantly affected by Zn, whereas
Zn transport was greatly inhibited by Cd. This finding implies that
Cd accumulation in this species is mediated at least partially by
Zn transporters which, however, seem to have higher affinity for
Cd than for Zn. Collectively, our results primarily indicate that A.
carambola has developed noteworthy physiological mechanisms to
accumulate Cd at a high level.
Acknowledgements
We thank Professor Alan J.M. Baker of the University of Mel-
bourne, Professor Hans Lambers of the University of Western
Australia and the two reviewers for their help in improving the
final version of this manuscript. This work was partially supported
by the International Foundation for Science, Stockholm, Sweden,
through a grant to Jin-tian Li (No. C/4785-1). We are also grate-
ful for financial support from the Science and Technology Planning
Project of Guangdong Province (2009B030802005) and the China
Postdoctoral Science Foundation (No. 20080440793).
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