Minerals Engineering 20 (2007) 188–190

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Technical note

Accumulation and uptake of manganese in a hyperaccumulator

Phytolacca americana
a,*
Yuan Min , Tie boqing b, Tang Meizhen c, Isao Aoyama d

a
State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, Shanghai 200092, China
b
College of Resources and Environment, Hunan Agricultural University, Changsha 410128, China
c
College of Life Science, Qufu Normal University, Qufu 273165, China
d
Research Institute for Bioresources, Okayama University, Kurashiki 710-0046, Japan

Received 10 March 2006; accepted 20 June 2006

Available online 28 August 2006

Abstract

Phytolacca americana (pokeweed) has been found to be a new manganese hyperaccumulator plant by means of field surveys on Mn-
rich soils and by Hydroponics experiments. This species not only has remarkable tolerance to Mn but also has extraordinary uptake and
accumulation capacity for this element. The maximum Mn concentration in the leaf dry matter was 8000 mg/g on Xiangtan Mn tailings
wastelands, with a mean of 6490 mg/kg. The species was characterized by a high translocation factor (>10.76). Under nutrient solution
culture conditions, manganese concentration in the shoots increased with increasing external Mn levels. The maximum concentration of
Mn in leaves reached 47.06 g/kg at 50 mmol/L Mn. This species provides a new plant resource for exploring the mechanism of Mn hyper-
accumulation and has potential for use in the phytoremediation of Mn contaminated soils.
Ó 2006 Elsevier Ltd. All rights reserved.

Keywords: Manganese; Hyperaccumulator; Phytolacca americana; Phytoremediation

1. Introduction
Manganese is an essential micronutrient and an activa-
tor of a number of enzymes involved in the tricarboxylic
acid cycle. However, exposure to excess manganese results
in manganese toxicity, including Parkinson-like symptoms,
and abnormalities of the reproductive system and the
immune system (Erikson and Aschner, 2003; Mark et al.,
2005). Phytoremediation is defined as the ‘‘use of plants
and their associated root-bound microbial communities
to remove, contain, degrade or render harmless environ-
mental contaminants’’ (Baker and Brooks, 1989). Approx-
imately 13 manganese hyperaccumulating plants are
currently known (Reeves and Baker, 2000).
The aim of this study was to determine the potential use
of Phytolacca americana as manganese hyperaccumultor
*
Corresponding author. Tel./fax: +86 21 6598 5662.
E-mail address: ym791114@yahoo.com.cn (Y. Min).
plant. Both plant and soil samples have been analyzed
for Mn at Xiangtan Mn tailings wastelands, Hunan Prov-
ince, southern China, and more detailed experiments have
been carried out on this species under controlled conditions
to investigate various aspects of its Mn accumulating
ability.
2. Materials and methods
2.1. Site situation
The manganese mine is located in Xiangtan country,
Hunan Province, southern China (27°40 0 –28°05 0 N,
112°37 0 –112°48 0 E), which has a subtropical continental cli-
mate with an annual average temperature of 17.4 °C, aver-
age rainfall of 1431.4 mm and average evaporation
capacity of 1321.7 mm. The soil pH value ranges from
7.46 to 7.65. The soil contained 117,830 mg/kg (average
value) of total Mn, 732.86 mg/kg (average value) of

0892-6875/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.mineng.2006.06.003
 Y. Min et al. / Minerals Engineering 20 (2007) 188–190 189

effective Mn, 765 lg/g of total nitrogen, 3430 lg/g of total 3. Results
phosphorus and 14.8% of clay (<0.01 mm) (Table 2).
3.1. The hyperaccumulating Mn characteristics
2.2. Plant and soil sampling of P. americana

According to the topography, plant and soil samples
were collected. The identification of plant species was made
according to Chinese weeds will (Li, 1998). The soil sam-
ples from corresponding top layer (depth for 0–30 cm) in
which plants grown were collected for element analysis.
2.3. Hydroponics culture
Hoagland’s nutrient solution was used to cultivate the
plant (Hoagland and Arnon, 1950), P. americana obtained
from the mining site. Two uniform plants for each treat-
ment group were selected and exposed to seven different
levels of Mn in solution: 0.1 (CK), 1, 2.5, 5, 10, 25 and
50 mmol/L Mn, added as MnSO4 Æ 7H2O. Each treatment
was replicated three times. The solutions were aerated for
24 h a day, adjusted daily to pH 5.8, and renewed every
four days. After 12 days treatment plants were harvested.
All plant samples were separated into roots, stems and
leaves. They were evaporated 30 min under 105 °C, and
then dried to constant weight at 70 °C. Fresh weight
(FW) and dry weight (DW) of roots, stems and leaves were
recorded. The samples of dried plant materials for Mn
analysis was ground.
2.4. Plant and soil sample analysis
P. americana often grew in dense clusters on the manga-
nese mine. The Mn concentration in the shoots was invari-
ably greater than that in roots, characteristic of
hyperaccumulators (Baker, 1981). In particular, the maxi-
mum Mn concentration in the leaf dry matter was
8000 mg/kg, and the maximum Mn concentration in the
root dry matter was 544 mg/kg. The corresponding soils
contained total Mn ranged from 93,575 to 141,775 mg/kg
(Table 1). The maximum ratio of Mn concentration in
leaves to that in roots was 15.56. However, the ratio of
Mn concentrations in leaves to that in soils ranged from
0.046 to 0.066.
3.2. Growth responses of P. americana to different Mn supply
levels
P. americana grew normally at Mn concentration below
10 mmol/L during the period of treatment, the biomass of
P. americana was increased with increasing of Mn concen-
tration. Its shoot dry weight had a sharp increases by 21%
compared with the control at 10 mmol/L (Table 2). When
Mn concentration was at 25 and 50 mmol/L, for 6 days, P.
americana presented yellow, wrinkle, curly and even wither,
and the dry weights of shoot decreased to 91% and 81%,
respectively (Table 2).

The available Mn was extracted with NH4OAc and
C6H4(OH)2 solution (Shi and Bao, 1981). The plant and
soil samples were digested with HNO3–HClO4, then to
determine its Mn concentration by using AAS method
(AA-646 Japan).
2.5. Statistical analysis
Analysis of variance was performed using DPS. Data
was expressed as means with standard error attached. A
test of equality of averages (LSD) was also applied.
3.3. Manganese accumulation and distribution
in P. americana
The concentrations of Mn in leaves, stems and roots
increased progressively with increase in Mn levels
(Fig. 1). The maximum concentration of Mn in leaves
reached 47.06 g/kg at 50 mmol/L Mn. In the present study,
there are remarkable differences in the distribution of Mn
both in shoots and roots of P. americana. Mn concentra-
tion in the shoots takes account for 86–95% of the whole
that plant absorbed.

Table 1
Concentration and mineralogical forms of Mn in soils and Mn bioconcentration and translocation of P. americana grown on Xiangtan manganese tailings
wastelands, Southern China
No. Mn in soils (mg/kg) Mn in plant tissues (mg/kg) L/Sa L/Rb S/Rc
Total Mn Exchangeable Mn Reducible Mn Effective Mn Leaf Stem Fruit Root
1 123,000 23 16,230 834.5 7180 620 730 542 0.058 13.25 1.14
2 120,850 31 17,516 906.8 8000 648 712 514 0.066 15.56 1.26
3 141,775 18 16,485 842.25 6980 578 654 521 0.049 13.39 1.11
4 93,575 13 11,654 595.7 5220 523 610 485 0.056 10.76 1.08
5 112,025 15 9865 508.28 5160 484 597 457 0.046 11.29 1.06
6 115,755 16 13,872 709.6 6400 560 640 544 0.055 11.76 1.03
a
Fraction of Mn concentrations in leaves to that in soils.
b
Fraction of Mn concentrations in leaves to that in roots.
c
Fraction of Mn concentrations in stems to that in roots.
190 Y. Min et al. / Minerals Engineering 20 (2007) 188–190

Table 2
The biomass and Mn concentration of P. americana grown in nutrient solution
Treatment (mmol/L) Dry weighta (g per plant) Mn contenta (mg per plant)
Leaf Stem Root Leaf Stem Root
CK 1.870 ± 0.064 0.429 ± 0.045 0.460 ± 0.048 1.152 ± 0.038 0.206 ± 0.013 0.150 ± 0.005
1 1.980 ± 0.073 0.440 ± 0.052 0.473 ± 0.055 10.043 ± 0.535 1.625 ± 0.295 1.270 ± 0.014
2.5 2.058 ± 0.088 0.453 ± 0.055 0.495 ± 0.061 16.252 ± 0.654 2.385 ± 0.350 2.303 ± 0.033
5 2.137 ± 0.110 0.482 ± 0.060 0.508 ± 0.063 19.293 ± 0.783 3.536 ± 0.415 3.211 ± 0.125
10 2.292 ± 0.135 0.501 ± 0.065 0.643 ± 0.071 26.954 ± 1.193 4.840 ± 0.493 5.365 ± 0.183
25 1.678 ± 0.053 0.415 ± 0.040 0.394 ± 0.045 66.348 ± 2.030 7.848 ± 0.705 4.275 ± 0.145
50 1.469 ± 0.045 0.403 ± 0.038 0.325 ± 0.042 69.131 ± 2.393 11.860 ± 0.898 4.117 ± 0.132
LSD0.05 0.152 0.085 0.097 2.352 0.937 0.156
LSD0.01 0.211 0.117 0.134 3.264 1.301 0.216
a
Means ± SE, n = 3.

50000 physiology and biochemical mechanisms of Mn hyperaccu-

root
45000 mulation.
Mn concentration(mg/kg)

stem
40000
leaf
35000
5. Conclusions
30000
25000
20000 Results from the present field surveys and the hydropon-
15000 ics trials revealed that P. americana is a manganese hyper-
10000 accumulator plant. Therefore, the mechanisms of Mn
5000 reduction, detoxification, and association with other ele-
0
CK 1 2.5 5 10 25 50 ments in Phytolacca americana L are the key objectives
Mn treatment(mmol/L) of our on-going research.
Fig. 1. Concentration of Mn in the tissues of P. americana.
Acknowledgement

4. Discussion The work was supported by the Cooperative Toyota’s

It is reported that a manganese hyperaccumulator
should have concentrations in excess of 1000 mg Mn
kg1 in its’ aboveground biomass (Baker and Brooks,
1989). In addition, its transfer factor should be greater than
one. However, normal concentrations of manganese in
plant dry matter fall within a rather wide range of 20–
500 mg/kg on normal soils (Reeves and Baker, 2000). In
the present study, the leaves of P. americana grown on
the Mn tailings wasteland contained as much as 8000 mg/
kg with a mean of 6490 mg/kg Mn (Table 1). The species
was characterized by a high translocation factor (>10.76).
And the maximum Mn concentration in the leaf dry matter
reached 47.06 g/kg under hydroponics culture conditions
(Fig. 1). Sang (2002) suggest that PAP-H is constitutively
produced by P. americana as a host defense protein, and
secreted into the rhizosphere as a barrier against soil-borne
microbe infection. The secretion of PAP-H is regulated in
response to endo- and exogenous stresses. In considering
the feasibility of phytoextraction, P. americana is a peren-
nial herb with a broad ecological amplitude, and it has
the characteristics of a fast growth rate, large biomass,
abundant seed production, and it would be easy to harvest
mechanically (Li, 1998). As can be seen from above discus-
sion, these traits of P. americana should make it a valuable
and convenient new plant material for further studies of the
fund projects of China and Japan (Toyota Fund D01-B3-
010).
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