Chemosphere 78 (2010) 1136–1141

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Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Cadmium accumulation and distribution in populations of Phytolacca americana

L. and the role of transpiration
Xiaoqing Liu a, Kejian Peng b, Aiguo Wang a, Chunlan Lian c, Zhenguo Shen a,*
a
College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, People’s Republic of China
b
Hunan Research Academy of Environmental Science, Changsha 410004, People’s Republic of China
c
Asian Natural Environmental Science Center, The University of Tokyo, 1-1-8 Midori-cho, Nishitokyoshi, Tokyo 188-0002, Japan

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

Article history: The concentrations of heavy metals in Phytolacca americana L. and corresponding soil samples from three
Received 7 October 2009 contaminated sites and an uncontaminated site were studied. Hydroponic experiments were also con-
Received in revised form 11 December 2009 ducted to investigate the Cd uptake ability and mechanism of P. americana. The field results showed that
Accepted 13 December 2009
the average Cd concentration was 42 mg kg1 in P. americana leaves, with the highest concentration of
Available online 13 January 2010
402 mg kg1 found at Datianwan. A significant relationship was observed between the concentrations
of Cd in leaves and those of corresponding soils on a logarithmic scale. Under laboratory hydroponic con-
Keywords:
ditions, the maximum Cd concentration in aerial tissues of P. americana was 637 mg kg1, under treat-
Phytolacca americana
Contaminated soil
ment with 100 lM Cd. The population from the uncontaminated site (Zijinshan) also had a remarkable
Cadmium accumulation ability to accumulate Cd in shoots to concentrations well in excess of 100 lM in the hydroponic exper-
Transpiration iment, similar to the population from contaminated site, suggesting that Cd accumulation is a constitu-
Abscisic acid tive trait of P. americana. In the presence of 100 lM Cd, the addition of polyethylene glycol decreased leaf
Populations transpiration, the shoot Cd concentration, and the shoot/root Cd concentration ratio. There was a signif-
icantly positive relationship between the shoot Cd concentration and the leaf transpiration of P. ameri-
cana. A similar significant positive correlation was also obtained between the shoot/root Cd
concentration and leaf transpiration. Moreover, pretreatment with 5 lM abscisic acid or 5 lM HgCl2 sig-
nificantly decreased the Cd concentration in P. americana shoots. These results suggest that transpiration
has an important role in Cd accumulation in shoots of P. americana.
Ó 2009 Elsevier Ltd. All rights reserved.

1. Introduction
Heavy metal contamination has received considerable public
attention over the last few decades. Cadmium is a highly toxic non-
essential element of particular concern to human health as it can
be readily absorbed by plant roots and be concentrated by many
cereals, potatoes, vegetables, and fruits (Wagner, 1993). There is
an urgent need for efficient and adaptable methods to remediate
soils contaminated with heavy metals and other contaminants.
Environmental remediation using the ability of endemic plants to
accumulate heavy metals has been proposed, termed phytoextrac-
tion; the phytoextraction of Cd from contaminated soils is indi-
cated in some cases (McGrath, 1998).
It is well recognized that the success of any phytoextraction
technique depends upon suitable plant species that can accumu-
late high concentrations of heavy metals and produce sufficient
biomass. The metal concentrations in plant shoots are generally
more important than the total biomass yield (McGrath, 1998; Shen
* Corresponding author. Tel.: +86 25 84396391; fax: +86 25 84396542.
E-mail address: zgshen@njau.edu.cn (Z. Shen).
et al., 2002), and the use of hyperaccumulators is essential for the
phytoextraction of heavy metals from polluted soils. Hyperaccu-
mulator plants are able to accumulate high amounts of heavy met-
als in their tissues, at concentrations 10–100 times higher than
those tolerated by crop plants (Baker and Brooks, 1989; Brooks,
1998). Over 450 plant species have been identified as hyperaccu-
mulators of trace metals (Zn, Ni, Mn, Cu, Co, and Cd), metalloids
(As), and nonmetals (Se) (Baker and Brooks, 1989; Verbruggen
et al., 2009). Hyperaccumulation of Cd in higher plants is a rare
phenomenon. Few plant Cd hyperaccumulators have been identi-
fied that can accumulate over 100 mg Cd kg1 in aerial parts. Some
European populations of Thlaspi caerulescens, a well-known Zn
hyperaccumulator plant, have been reported to accumulate signif-
icantly high concentrations of Cd in their aerial parts, reaching
3000 mg kg1 (Lombi et al., 2000). Another possible Cd hyperaccu-
mulator is Arabidopsis halleri (Brooks, 1998), which is able to accu-
mulate up to 6000 mg Cd kg1 in shoots under hydroponic culture
conditions (Küpper et al., 2000), and more than 100 mg Cd kg1
under natural conditions (Bert et al., 2002). Recently, Viola bao-
shanensis (Liu et al., 2004), Salsola kali (de la Rosa et al., 2004), Se-
dum alfredii (Yang et al., 2004), and Thlaspi praecox (Vogel-Mikuš

0045-6535/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2009.12.030
 X. Liu et al. / Chemosphere 78 (2010) 1136–1141 1137

et al., 2005, 2008) have also been reported to hyperaccumulate Cd quent hydroponic experiments were conducted to investigate the
from polluted soils. relationship between transpiration and the accumulation of Cd in
The mechanisms of Cd hyperaccumulation have been studied shoots.
extensively, predominantly in T. caerulescens (Lombi et al., 2001,
2002; Zhao et al., 2002b; Roosen et al., 2003). Physiological studies 2. Materials and methods
have provided strong evidence that multiple uptake systems are
involved in the uptake of Cd by roots. The Vmax for Cd uptake is 2.1. Plant and soil sampling
approximately five times higher in the Ganges ecotype than in
the Prayon ecotype of T. caerulescens (Lombi et al., 2001). The Cd The field investigations were carried out in 2003 and 2007.
influx kinetics for Ganges showed a saturable component at low Leaves of P. americana and soil samples were collected from three
Cd concentrations, which is very weak in Prayon. The uptake of contaminated sites and one uncontaminated site: one wasteland
Cd by the Ganges ecotype is not inhibited by La, Zn, Cu, Co, Mn, around the Xiangxi Zn and Mn smelter in Jishou city, Hunan Prov-
Ni, or Fe(II), nor by increasing the Ca concentration (Zhao et al., ince (109°45.10 E, 28°17.80 N); two Cu mine tailing ruins in Tongling
2002b). These results suggest the existence of a specific transport city, Anhui Province (117°49.10 –117°53.50 E, 30°55.60 N); and one
system with high affinity for Cd in the Ganges ecotype. In contrast, uncontaminated site in Nanjing city, Jiangsu Province
Cd uptake by the Prayon ecotype may be mediated partly via Ca (118°50.10 E, 32°03.50 N). At least 10 individual leaves from each
channels or transporters for Zn and Mn. Roosen et al. (2003) sug- plant were collected and mixed to give a representative sample.
gested that Cd is taken up predominantly by a high-affinity uptake The soils in which the sampled plants were growing were also col-
system for Fe in the Ganges ecotype. However, Plaza et al. (2007) lected for Cd analysis. The soil sampling depth was about 0–20 cm.
reported that although TcIRT1 (T. caerulescens iron-regulated trans-
porter) may be involved in Cd hyperaccumulation in the Ganges
2.2. Hydroponic experiments
ecotype, the transporter expressed in yeast does not show en-
hanced ability to transport Cd.
Seeds of two populations of P. americana were collected from
Water and ions can also enter roots directly via a low-resistance
Zijinshan, Nanjing city, Jiangsu Province, and from Datianwan in
apoplastic pathway (Ma and Peterson, 2003). However, the non-
Xiangxi, Hunan Province, China. The seeds were sown in vermicu-
selective apoplastic bypass flow of water and ions into the root
lite and germinated in a growth chamber with a photoperiod of
stele may be blocked by the presence of Casparian bands in radial
12 h light/12 h dark and day/night temperatures of 25/20 °C for
and transverse walls of the endodermis and exodermis. It has been
15 d. The seedlings were transferred to a vessel containing 2 L of
shown that in healthy roots, apoplastic bypass to shoots is mini-
Hoagland’s nutrient solution with the pH adjusted to 5.6 ± 0.1 with
mal, but increases if the plant is subjected to anaerobic stress or
HCl or NaOH. The nutrient solution was renewed every 2 d. Four-
physical damage (Hanson et al., 1985; Moon et al., 1986). White
teen days later, the seedlings were treated with 0, 10, or 100 lM
et al. (2002) suggested a significant role of the apoplastic pathway
Cd in Experiment 1, supplied as CdCl2 in Hoagland’s solution. Each
in xylem loading of Zn in T. caerulescens. It was noted that Caspar-
treatment was replicated in three different vessels, with each ves-
ian bands are not totally impermeable to water, or even to ions
sel containing five plants. After a further 10 d of growth, the plants
(Ranathunge et al., 2005). However, Xing et al. (2008) reported that
were harvested, and the shoots and roots were separated, washed
the translocation efficiency of Cd and Zn did not correlate with the
thoroughly with tap water, rinsed with deionized water, blotted
apoplastic bypass flow measured with a membrane-impermeable
dry with tissue paper, weighed, dried at 80 °C for 24 h, and
fluorescent dye.
weighed again. Experiments 2–4 were conducted to study the role
The effect of transpiration on Cd accumulation in the shoots has
of transpiration in Cd accumulation in shoots of P. americana. In
been investigated in several species (Florijn and Van Beusichem,
Experiments 2 and 3, seedlings were pretreated with 5 lM ABA
1993; Salt et al., 1995; Hsu and Kao, 2003; Ingwersen and Streck,
(abscisic acid) or HgCl2 for 1 d, respectively, and then 10 or
2005; Salah and Barrington, 2006; Van der Vliet et al., 2007). Salt
100 lM Cd were added. After a further 2 d of growth, net photo-
et al. (1995) suggested that Cd accumulation in leaves is mainly
synthesis, transpiration, and stomatal conductance were measured
driven by transpiration, and higher transpiration generally leads
using a Li-6400 Portable Photosynthesis System (Li-Cor Inc., USA).
to higher shoot Cd levels (Salah and Barrington, 2006). The differ-
The plants were then harvested for analysis of Cd concentrations.
ential accumulation of Cd by different cultivars of durum wheat
In Experiment 4, 14-d-old seedlings were grown in solutions with
could be partially attributed to the effect of Cd on transpiration
0–10% polyethylene glycol 4000 (PEG) (n = 22) in the presence of
(Van der Vliet et al., 2007). In maize inbred lines, however, no rela-
100 lM Cd for 1 d. Leaf transpiration rates were measured with
tionship was found between the shoot Cd concentration and the
the Li-6400 Portable Photosynthesis System, and the plants were
transpiration coefficient (Florijn and Van Beusichem, 1993). These
then harvested for analysis of Cd concentrations.
different results may be due to different experimental conditions
as well as differences between species.
Phytolacca americana is a perennial weed that occurs world- 2.3. Sample preparation and analysis
wide. According to our investigations, it is often found in disturbed
habitats, particularly in contaminated areas of South China. Previ- Soil samples were air-dried at room temperature for 2 week,
ous results have shown that P. americana can accumulate large ground into a fine powder, passed through a 0.5-mm nylon sieve,
amounts of heavy metals in its aerial tissues, especially Cd and and digested with HF:HNO3:HClO4 (4:1:1, v/v). The dried plant
Mn (Peng et al., 2006, 2008), with concentrations reaching samples were ground and completely digested with a solution of
10 700 and 408 mg kg1 DW, respectively, in leaves of plants ex- extra-pure HNO3 and HClO4 (87:13, v/v). The Cd concentrations
posed to 5000 lM Mn with 50 lM Cd (Peng et al., 2008). In the were determined using an atomic absorption spectrophotometer
presence of Cd, adding Mn to the solution significantly improved (model TAS-986, Purkinje General Co. Ltd., Beijing, China).
plant growth, and reduced the Cd concentrations in all organs of
the plant. However, the mechanisms of metal tolerance and accu- 2.4. Statistical analysis
mulation in P. americana are unclear.
The present work was undertaken to further study the Cd-accu- Data were analyzed using SPSS ver. 13.0 (Statistical Package for
mulating ability of P. americana growing in polluted areas. Subse- Social Science for Windows, ver. 13.0) for the ANOVA test. Signifi-
1138 X. Liu et al. / Chemosphere 78 (2010) 1136–1141

cant differences among means were determined by the LSD at 1000

Cd content in leaf (mg kg )

P < 0.05.

-1
100
3. Results

3.1. Cd accumulation by plants at contaminated and uncontaminated 10

sites

Table 1 shows the range and mean of Cd concentrations in 1 R2 = 0.7697

leaves of P. americana from contaminated sites (Datianwan, Shizi- n=116
shan, and Tongguanshan) and the uncontaminated site (Zijinshan),
0.1
and the corresponding soils. The leaf Cd concentrations ranged 0.1 1 10 100 1000 10000
from undetectable to 402 mg Cd kg1, showing the extraordinary Concentration of total Cd in soil (mg kg )
-1
ability of P. americana to accumulate Cd in leaves. Leaves of plants
grown at the Datianwan site had higher Cd concentrations than Fig. 1. Relationship between concentrations (mg kg1) of Cd in leaves of P.
those from other sites. The average Cd concentrations were 151, americana and in the corresponding soils.
18, 15, and 1.2 mg kg1 in leaves of P. americana grown at Datian-
wan, Shizishan, Tongguanshan, and Zijinshan, respectively.
Table 2
P. americana can grow in soil containing up to Dry weights (mg plant1) of P. americana exposed to Cd for 10 d under hydroponic
1083 mg Cd kg1 DW. On a logarithmic scale, the concentration culture conditions.
of Cd in leaves was significantly correlated with the soil Cd concen-
Cd treatment (lM) Shoots Roots
tration (Fig. 1). The average leaf bioaccumulation factor (plant/soil
concentration ratio) for Cd was 0.93, ranging from 0.06 to 9.7, and Datianwan Zijinshan Datianwan Zijinshan

29% of the plants had a bioaccumulation factor greater than 1.0. 0 347 351 91 94
The bioaccumulation factor for Cd tended to decrease with increas- 10 290 288 96 94
100 200 190 55 63
ing soil Cd concentration (Table 1).
ANOVA F ratio
Population 0.0750NS 0.8170NS
3.2. Plant growth and Cd uptake under hydroponic culture conditions Cd 96.06*** 52.64***
Population  Cd 0.2030NS 0.9810NS
Error df 10 10
The growth and Cd uptake of two P. americana populations, one
LSD (P < 0.05) 35.25 12.30
from a contaminated site (Datianwan) and the other from the
uncontaminated site (Zijinshan), were compared in hydroponic NS: not significant.
***
P < 0.001.
culture. The effects of Cd treatment were generally consistent in
both populations: 10 and 100 lM Cd caused leaf chlorosis and pro-
duced a significant decrease in the shoot dry weight (Table 2).
Table 3
Treatment with 10 lM Cd had no significant effect on the root Concentrations of Cd (mg kg1) in P. americana exposed to Cd for 10 d under
dry weight. The Cd concentrations in shoots and roots of both pop- hydroponic culture conditions.
ulations increased with increasing Cd concentration in the nutrient
Cd treatment (lM) Shoots Roots
solution (Table 3). The highest concentration of Cd in shoots was
637 mg Cd kg1 dry mass in the 100 lM Cd treatment. In all Cd Datianwan Zijinshan Datianwan Zijinshan

treatments, the roots of both populations accumulated more Cd 0 nd nd nd nd

than the shoots. No significant difference in the Cd concentrations 10 116 130 493 481
100 637 614 1185 1171
in shoots and roots were found between the two populations.
ANOVA F ratio
Population 0.0760NS 0.2570NS
3.3. Effect of ABA and HgCl2 pretreatment on growth and Cd Cd 1058*** 1578***
accumulation Population  Cd 0.8090NS 0.0650NS
Error df 10 10
LSD (P < 0.05) 45.42 66.37
Compared with the 10 lM Cd treatment, treatment with
100 lM Cd for 2 d significantly decreased the transpiration and NS: not significant; nd: not detectable (<the detection limit, Cd: 0.1 mg kg1).
***
P < 0.001.
stomatal conductance of P. americana, but had no significant effect

Table 1
Concentrations of Cd and bioaccumulation factors in P. americana collected from
contaminated and uncontaminated sites.
on the net photosynthesis in leaves. Roots of P. americana were
Site Year No Cd (mg kg1) range (a.m) BF range pretreated with 5 lM ABA for 1 d before exposure to solutions con-
(a.m)
Leaf Soil taining 10 or 100 lM Cd. Compared with plants that received no
Datianwan 2003 16 5–402 (109) 8–1083 (187) 0.12–5.0 (1.2) pretreatment, plants pretreated with 5 lM ABA showed signifi-
Datianwan 2007 9 51–388 51–1037 0.25–3.0 cantly decreased transpiration and stomatal conductance and
(226) (585) (0.66) slightly decreased net photosynthesis (Table 4). ABA pretreatment
Zijinshan 2007 30 nd–2 (1.2) 0.2–5 (3) 0.06–9.7
also significantly decreased the Cd concentrations in P. americana
(0.71)
Tongguanshan 2007 32 4–39 (15) 8–50 (18) 0.16–1.9
shoots and roots (Table 4).
(0.90) Pretreatment (1 d) with 5 lM HgCl2, which is known to inhibit
Shizishan 2007 30 7–28 (18) 13–35 (17) 0.24–4.9 (1.1) bulk water transport through aquaporins, also decreased the Cd
nd: not detectable (<the detection limit, Cd: 0.1 mg kg1); a.m: arithmetic mean;
accumulation in shoots and roots of P. americana (Table 5). The de-
BF, bioaccumulation factor = metal concentration in leaf/metal concentration in crease in Cd concentration was more pronounced in shoots than in
soil. roots.
 X. Liu et al. / Chemosphere 78 (2010) 1136–1141 1139

Table 4
Effects of ABA on the net photosynthesis, transpiration, stomatal conductance, and Cd concentrations of P. americana in the presence of 10 or 100 lM Cd.

Pretreatment ABA Treatment CdCl2 Net photosynthetic rate (lmol CO2 Transpiration rate (mmol H2O Stomatal conductance (mol H2O Cd (mg kg1)
(lM) (lM) m2 s1) m2 s1) m2 s1)
Shoots Roots
0 10 8.8 3.1 0.12 40 501
5 10 7.0 1.4 0.05 26 255
0 100 8.1 1.9 0.08 285 2020
5 100 5.5 0.9 0.03 166 1719
ANOVA F ratio
Cd 3.664NS 8.290* 6.029* 461.1*** 4576***
ABA 13.84* 18.89** 25.80** 55.93 153.2***
***

Cd  ABA 0.3760NS 1.357NS 0.4540NS 34.04** 1.589NS

Error df 6 6 6 6 6
LSD (P < 0.05) 2.031 1.038 0.0380 30.96 76.31

NS: not significant.

***
P < 0.001.
**
P < 0.01.
*
P < 0.05.

3.4. Effect of transpiration on Cd accumulation and distribution the ability of P. americana to accumulate Cd in its leaves is extraor-
dinary compared to other plant species in Datianwan (Peng et al.,
To collect information on the effect of transpiration on Cd accu- 2006). These results suggest that the Cd resistance of P. americana
mulation in P. americana shoots, plants were exposed for 1 d to could be sufficient to allow growth on contaminated soils, and
nutrient solutions containing different concentrations of PEG in would facilitate the colonization of metal-contaminated sites from
the presence of 100 lM Cd, and the leaf transpiration and Cd con- adjacent populations. This may also be the basis for the universal
centrations in shoots and roots were measured. The concentrations growth of this species on metal-contaminated sites.
of Cd in P. americana shoots and roots increased significantly with The bioaccumulation factor (the plant/soil concentration ratio)
increasing leaf transpiration (Fig. 2). A significant increase in the is more important than the shoot concentrations when considering
shoot/root ratio of Cd concentrations occurred with increasing leaf the potential phytoremediation of a given species (Zhao et al.,
transpiration, indicating that less Cd was transported from roots to 2003). In metal-accumulating species, the bioaccumulation factor
shoots when leaf transpiration was inhibited. is often greater than 1, whereas in metal-excluder species the fac-
tor is typically below 1 (Baker, 1981). In P. americana leaves, the Cd
bioaccumulation factors based on the total Cd soil concentrations
4. Discussion ranged from 0.06 to 9.7. If hyperaccumulation is defined as having
a bioaccumulation factor of greater than 1 (Baker, 1981), only
Among plant species recognized as hyperaccumulators, only about 29% of the P. americana individuals met this criterion.
four show the capacity to hyperaccumulate Cd (Verbruggen et al., Although the Cd bioaccumulation factors were lower than those
2009). P. americana is widely distributed in uncontaminated and reported by Zhao et al. (2003) for T. caerulescens, P. americana has
contaminated areas of South China. In the present field survey, P. great potential in phytoextraction from Cd-contaminated soils be-
americana demonstrated high adaptability to a wide range of soil cause of its rapid growth and high above-ground biomass. Field
Cd concentrations, and accumulated high Cd concentrations in its observations indicated that P. americana attained a height of over
aerial parts. 60% of the plants from the Datianwan site contained 1.8 m with leaf Cd concentrations >100 mg Cd kg1. Moreover, this
>100 mg Cd kg1 DW in leaves, the threshold value commonly species can re-grow shoots from its fleshy taproot after above-
used to define Cd hyperaccumulation in natural habitats (Baker ground parts are harvested, eliminating the need for seed sowing.
and Brooks, 1989). The normal concentration of Cd in plants ranges Finally, this species can tolerate high levels of Cu, Pb, and Zn in
from 0.05 to 0.2 mg kg1 (Kabata-Pendias and Pendias, 1992), and soils (Peng et al., 2006), and would thus be suitable for use in mul-
ti-metal-contaminated sites.
The P. americana metal concentrations strongly depend on the
Table 5 metal concentrations in the soil. It is noteworthy that the popula-
Effects of HgCl2 on the Cd concentrations of P. americana in the presence of 10 or tion from the uncontaminated site (Zijinshan) also had a remark-
100 lM Cd. able ability to accumulate Cd in shoots to concentrations well in
Pretreatment HgCl2 (lM) Treatment CdCl2 (lM) Cd (mg kg1) excess of 100 lM in the hydroponics experiment, similar to popu-
lation from other contaminated site. A similar result was observed
Shoots Roots
for another population from an uncontaminated site in Huaihua,
0 10 36 462 Hunan Province (unpublished data). Metal accumulation appears
5 10 6 111
0 100 175 1004
to be a constitutive trait at the species level, as found by Bert
5 100 74 851 et al. (2002) in metallicolous and nonmetallicolous populations
ANOVA F ratio of A. halleri that had the same ability to accumulate Zn and Cd,
Cd 188.4*** 563.6*** and by Zhao et al. (2002a) in the As hyperaccumulator Pteris vittata.
HgCl2 75.77*** 87.08***
However, the general response and Cd accumulation of T. caerules-
Cd  HgCl2 21.80** 13.34*
Error df 6 6 cens differed significantly between populations (Basic et al., 2006),
LSD (P < 0.05) 26.05 93.51 and three populations of S. alfredii exhibited population-specific
*** and metal-specific variations in Zn and Cd accumulation (Deng
P < 0.001.
**
P < 0.01. et al., 2007). In contrast to Zn, the Cd-accumulation abilities of
*
P < 0.05. the populations appeared to vary, as observed in T. caerulescens
1140
1000
a
Cd content in shoot (mg kg-1)
100
10
1
10000
b motion of Cd and water apoplastic flows by transpiration. Inhibit-
Cd content in root (mg kg )
-1
1000
100
10
1 uted to shoots of A. halleri. In contrast, it has been suggested that
1
c from the soil solution to plant tissues (Van der Vliet et al., 2007).
Cd content in shoot/root
0.1
0.01
0.1
Transpiration rate (mmol H 2O m-2 s-1)
Fig. 2. Relationships between the shoot Cd concentration (a), root Cd concentration
(b), and shoot/root ratio of Cd concentration (c) of P. americana and leaf
transpiration.
(Lombi et al., 2000) and A. halleri (Bert et al., 2002). There are few
systematic differences between different populations in soil and
plant Zn concentrations, whereas for Cd, there is a distinct differ-
ence between populations from France and those from other areas
in the shoot/soil metal relationship (Zhao et al., 2003).
The bioaccumulation factor for Cd tends to decrease with
increasing total soil Cd concentrations. A similar result was re-
ported in populations of T. caerulescens (Zhao et al., 2003) and T.
praecox (Vogel-Mikuš et al., 2005). The decrease in the bioaccumu-
lation factor may be due to the saturation of metal uptake and/or
the root-to-shoot transport when the internal metal concentration
is high (Zhao et al., 2003).
Little is known about the mechanisms of Cd accumulation in P.
americana. Cd accumulation in plant shoots appears to be con-
trolled by several processes, including root uptake, radial transport,
xylem loading, and translocation from the root to the shoot. Tran-
spiration is the driver for bulk water flow in these processes. In
wastewater irrigation areas, transpiration plays an important role
in Cd uptake by field crops (Ingwersen and Streck, 2005). Between
66% and 82% of the relationship between Cd concentration in crops
and in the soil solution can be explained by the amount of water
transpired. Salt et al. (1995) suggested that Cd translocation to
X. Liu et al. / Chemosphere 78 (2010) 1136–1141
the shoot of Brassica juncea is mainly driven by mass flow due to
transpiration. The present study suggests that transpiration might
be involved in Cd accumulation in P. americana shoots, based on
the observations that: (a) there was a significant positive relation-
ship between leaf transpiration and the shoot Cd concentration, (b)
pretreatment with ABA caused stomatal closure and significantly
decreased the transpiration rate and Cd concentrations in shoots,
and (c) pretreatment with the aquaporin inhibitor HgCl2 also sig-
R2 = 0.5827 nificantly decreased the shoot Cd concentration. Our result that
n=22 transpiration affects the accumulation of Cd in P. americana shoots
is consistent with observations from several other studies (Salt
et al., 1995; Kashem and Singh, 2002; Ingwersen and Streck,
2005; Salah and Barrington, 2006), and may be ascribed to the pro-
ing leaf transpiration decreases the translocation of Cd from roots
to shoots, as reported by Hsu and Kao (2003): exogenous applica-
tion of ABA reduced transpiration, decreased the Cd concentration,
and enhanced the Cd tolerance of a Cd-sensitive rice cultivar. The
exogenous application of an ABA biosynthesis inhibitor (fluridone)
decreased ABA accumulation, increased transpiration and the Cd
R2 = 0.706 concentration, and decreased the Cd tolerance of a Cd-tolerant rice
n=22 cultivar. Zhao et al. (2006) also found that pretreatment with ABA
dramatically decreased the proportion of 109Cd and 65Zn distrib-
more than one physiological process controls Cd translocation
Florijn and Van Beusichem (1993) found no relationship between
shoot Cd concentrations and the transpiration coefficient of maize,
and concluded that internal distribution rather than uptake is
mainly responsible for genotypic variations in maize shoot Cd con-
centrations. Further investigation of Cd uptake, translocation, and
storage in P. americana will be required using genetic and molecu-
lar techniques to achieve an understanding of the metal-accumula-
R2 = 0.4178 tion mechanisms of this species.
n=22
5. Conclusions
1 10
Results from a field survey showed that the Cd concentration in
P. americana leaves greatly depends on the soil Cd concentration.
Under laboratory hydroponic conditions, the population collected
from an uncontaminated site could accumulate Cd in shoots, sim-
ilar to populations from contaminated sites, suggesting that Cd
accumulation is a constitutive trait of P. americana. A significant
positive relationship was observed between the shoot Cd concen-
tration and the leaf transpiration rate of P. americana. Pretreatment
with 5 lM ABA or 5 lM HgCl2 also significantly decreased leaf
transpiration and the shoot Cd concentrations. The results demon-
strate that transpiration has an important role in Cd accumulation
in shoots of P. americana.
Acknowledgments
This study was supported by the National Special Program on
Water (2009ZX07212-001) and ‘‘863” Program (2007AA021404)
from the Ministry of Science and Technology of the People’s Repub-
lic of China, the 111 Project from the Ministry of Education of the
People’s Republic of China (No. B07030) and the key project from
the Science Foundation of Jiangsu Province (BK2006711-1).
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