全 文 ：5期
0 Introduction
【Research significance】P. vittata L.，the first-
known arsenic （As） hyperaccumulator （Ma et al .，
2001； Chen et al.，2002），is a relatively fast-growing
perennial fern. It can accumulate a huge biomass and
high concentrations of As in its above ground parts，
containing up to 5070 mg/kg As on a dry weight basis
（Chen et al.，2002）. The information on accumulation
of As and its distribution in pinnae of P. vittata L. was
limited. Research on this topic will further our under-
standing of the process of As absorption and transport
in P. vittata，and will provide information that will op-
timize collection of plant samples for phytoremediation
projects. Appropriate sample collection techniques will
Accumulation of arsenic and its distribution in pinnae of
hyperaccumulator Pteris vittata L.
WANGXue-li1，2*，WEIYan-yan1*，HEBing1，LEIMei2，GUMing-hua1**
（1College of Agriculture，Guangxi University，Nanning 530005，China；2Institute of Geographic Sciences and Natural
Resources Research，Chinese Academy of Sciences（CAS），Beijing 100101，China）
Abstract：【Objective】Chinese brake fern （Pteris vittata L） has been identified as an arsenic （As） hyperaccumula-
tor. The present study was conducted to obtain insight into accumulation of As and its distribution in pinnae of P. vittata
L.，which provide the basis to optimize the sampling method and evaluate the phytoremediation efficiency accurately.
【Method】Field sampling and laboratory analysis were used to research As accumulation and distribution in pinnae of hy-
peraccumulator P. vittata L. 【Result】As concentration in pinnae varied depending on their position along the frond. As
concentration in the middle pinnae accounted for 35% and 37% of the basal and top pinnae，respectively. Therefore，
there was a “U”-type distribution of As concentration along the frond. For As accumulation，based on biomass and As
concentration in the middle pinnae was 3.1 times and 2.8 times higher than those in the basal and top pinnae，showing a
“∩”-type distribution in the high-As P. vittata. 【Conclusion】As concentration in the middle pinnae is low，while As
accumulation was high，therefore，the basal，middle and top pinnae should be given consideration while evaluating the
phytoremediation efficiency in phytoremediation projects accurately.
Key words： P. vittata L.； arsenic； pinnae； accumulation； distribution
CLC number： X171.5 Document code：A Article：2095-1191（2015）05-0755-05
Received date：2015-04-05
Foundation item：National Natural Science Foundation（41071215）；Guangxi Natural Acience Foundation（2014GXNSFBA118223）
Biography：** for corresponding author，Gu Ming-h a（1962-），professor，inte esting in heavy metal pollution，E-mail：gumh@gxu.
edu.cn。 * Co-first author：WANG Xue-li（1982-） ，Ph. D.，interesting in heavy metal pollution and phytoremediation，
E-mail：wxl0524@126.com；WEI Yan-yan（1985-），Ph. D.，interesting in heavy metal pollution，E-mail： any nwei2008@
163.com
砷在超富集植物蜈蚣草羽片中的分布规律
王学礼1，2*，韦燕燕1*，何 冰1，雷 梅2，顾明华1**
（1广西大学 农学院，南宁 530005； 2中国科学院 地理科学与资源研究所，北京 100101）
摘要：【目的】探明砷在超富集植物蜈蚣草不同叶位羽片中的分布，为优化采样方法进而准确评估植物修复效率
提供依据。【方法】采用大田取样与室内分析相结合的方法，研究不同叶位的蜈蚣草羽片中砷分布的规律。【结果】蜈
蚣草不同叶位羽片中砷浓度差异较明显：中部羽片砷浓度分别是基部羽片和顶部羽片的35%和37%，由基部羽片到
顶部羽片砷浓度变化现先降低后升高的变化趋势，随着羽片叶位增加呈“U”型分布。蜈蚣草羽叶中部羽片砷富集量
是基部羽片的3.1倍，是顶部羽片的2.8倍，蜈蚣草羽叶砷富集量中部羽片较高，基部羽片和顶部羽片较低，呈倒“U”
型分布。【结论】中部羽片砷浓度较低，但砷累积量较大，因此通过采集羽片样品来评估蜈蚣草修复效率时要兼顾底
部羽片、中部羽片和顶部羽片。
关键词：蜈蚣草；砷；羽片；富集；分布
DOI：10.3969/j：issn.2095-1191.2015.5.755
南方农业学报 Journal of Southern Agriculture 2015，46（5）：755-759
ISSN 2095-1191；CODEN NNXAAB http：//www.nfnyxb.cn
南 方 农 业 学 报 46卷
allow accurate evaluation of phytoremediation efficien-
cy，and will help to determine the best planting density
for phytoremediation of As-contaminated soil. There-
fore，the aim of this study is to reveal accumulation of
As and its distribution in pinnae of P. vittata L. 【Re-
search progress】P. vittata had been proven to suc-
cessfully remediate As-contaminated soils （Chen et
al.，2007）. Previous studies reported that most amount
of As accumulated in the pinnae of P. vittata （Lombi
et al.，2002；Zhou et al.，2014； Ye et al.，2014）. Syn-
chrotron radiation -X -ray fluorescence spectrometry
analysis showed that the P. vittata root is able to
transport As from the cortex to the vascular tissues，
from which it was quickly transported from the xylem
of the frond rachis to the cortex and epidermis cells
（Huang et al.，2004a； Huang et al.，2011； Wan et
al.，2014）. Studies on chloroplast ultrastructure showed
that addition of high concentrations of As did not
damage chloroplast structure in new fronds，but had
severe negative effects on chloroplast membrane struc-
ture in mature secondary pinnae （Li et al.， 2005； Shen
et al.， 2014a，2014b）. The variation in As concentra-
tion among P. vittata pinnae of different ages could be
up to 36% （Ma et al.，2007），with the maximum As
concentrations in older fronds，and the minimum in
younger fronds （Tu and Ma ， 2005； Zhang et al.，
2012 ） . 【Research breakthrough point】There is no
report about the relationship between pinna phyllotaxy
of hyperaccumulator P. vittata L. and As distribution，
which is directly related to the representation of the
samples and then affect the accuracy of the evaluation
of phytoremediation efficiency. P. vittata plants were
collected from an As-phytoremediation field in Hunan
province in this study，and the differences in As dis-
tribution in pinnae between fronds with low and high
As concentrations were studied. 【Solving problems】
Field sampling and laboratory analysis were used to
research As accumulation and distribution in pinnae of
hyperaccumulator P. vittata L.，which will provide the
basis for reasonable collecting plant samples and then
evaluate the phytoremediation efficiency in phytore-
mediation projects accurately.
1 Materials and methods
1. 1 Sampling area
P. vittata plants were collected from an As-phy-
toremediation field in Hunan Province in December
2013，in central southern China. The As concentration
in the soil ranged from 36 to 98 mg/kg. The sample
collection area was located between a latitude of
24°53′ and 26°50′，and a longitude of 112°50′ E and
114°14′ E. There was a subtropical climate，with an-
nual average rainfall of 1500 mm in this area.
1. 2 Schematic diagram ofP.vitatpinnae on the
frond
Figure 1 showed a schematic diagram of P. vittata
pinnae on the frond. Pinnae were numbered from the
basal pinnae towards the top. The As concentration in
each pinna was defined as the average As concentra-
tionin the pinnae on the left and right sides of the
rachis. Similarly，the amount of As accumulation in
each pinna was defined as the average amount of As
accumulation in the pinnae on the left and right sides
of the rachis.
Taking the variability of data into account and for
the convenience of plotting，As concentration ration
was defined the ration of As concentration in a single
pinnae and the mean value of As concentration in the
opposite pinnae to study the pattern of As concentra-
tion along the frond. As enrichment amount ratio was
defined the ration of As accumulation in a single pin-
nae and the mean value of As enrichment in the oppo-
site pinnae to study the pattern of As accumulation a-
long the frond.
1. 3 Treatment of soil and plant samples
The frond was washed with distilled water，and
dried in the oven at 60 ℃ for 2 d. Soil samples were
digested with HNO3-H2O2，and plant samples were di-
gested with HNO3-HClO4 （5∶1） （USEPA3050B）. The
digested solutions were analyzed for As concentration
using an atomic fluorescence spectrometer （AFS -
2202，Haiguang Instrumental，Beijing，China）. Stan-
Fig.1 Schematic diagram of pinnae number in a frond of
P. vittata
Ci=1/2（CLi+CRi）
Ci：As concentration（mg/kg）of the i pinna；i：the number of the pinnaac-
cording to its position along the frond；CLi and CRi：As concentrations
（mg/kg）of the left and right pinnaeof the i pinna，respectively.
Mj=1/2（MLj+MRj）
Mj：Amount of As enrichment（g）in the j pinna；j：the number of the pin-
naaccording to its position on the frond；MLj and MRj：Amounts of As en-
richment（g）in the left and right pinnaeof the j pinna，respectively.
Journal of Southern Agriculture756· ·
5期 WANG et al.：Accumulation of arsenic and its distribution in pinnae of hyperaccumulator Pteris vittata L.
dard reference materials for plants （GBW-07603 and
GBW-08501），obtained from the China National Cen-
ter for Standard Reference Materials，were also treated
and analyzed in the same way，as used for the Quality
Control & Quality Assurance program. Statistical anal-
yses were conducted using SPSS statistical software
（SPSS Inc.，Chicago，U.S.A.，Release 11.0）. The sig-
nificance of differences in As concentrations among
different pinnae was tested using a LSD single-factor
analysis of variance with a significance level of 0.05.
2 Results and analysis
The As concentrations of 10 soil sampled obtained
from P. vittata plants grown field ranged from 36 to 98
mg/kg. The soil As concentration were ranked as fol-
lows：i（98 mg/kg）>d（90 mg/kg）>j（87 mg/kg）>h （71
mg/kg）>g （61 mg/kg）>b （54 mg/kg）>c （52 mg/kg）>f
（45 mg/kg）>e（43 mg/kg）>a（36 mg/kg）. The sampled
P. vittata plants did not show abnormal physiological
or morphological characteristics.
2. 1 As concentration in pinnae
T here were differences in As distribution in pin-
nae between fronds with low and high As concentra-
tions （Fig.2） . In fronds with low As concentrations
（255-590 mg/kg）， the middle of pinnae contained
lower As concentrations than other parts of pinnae，
showing the trend of As concentration decreased from
the basal to the middle pinnae，then increased towards
the top pinnae. The dynamic equation of heavy metals
parabolic diffusion equation was used to simulate the
data of As concentration（Xie et al.，2008），data fitting
effect was good，the correlation coefficient R2 in the
dynamic equation of low -As fronds was 0.98，fitting
the equation：Y=1.05 -0.05X +0.003X2. However，in
fronds with high concentrations of As （1040-1367
mg/kg），the As concentration decreased gradually from
the basal pinnae upwards，with only a slight increase
in As concentration in the top pinnae，and the correla-
tion coefficient R2 in the dynamic equation of high-As
fronds was 0.99，fitting the equation： Y=1.05-0.05X+
0.002X2. Therefore，the As concentration in basal pin-
nae was similar to that in the top pinnae in the low-
As fronds，but was higher in the basal pinnae than the
top pinnae in the high-As fronds （Fig.2）.
As concentrations in the pinnae were examined.
The As concentration was lower in the middle pinnae
and higher in the top and basal pinnae. As concentra-
tions in the middle pinnae of low P. vittata was 202
mg/kg，and As concentrations in the top and basal
pinnae was 577 mg/kg and 546 mg/kg，respectively.
The As concentration in the middle pinnae accounted
for 35% and 37% of the basal and top pinnae. There-
fore， there was a “U”-type distribution of As con-
centration along the frond.
Fig. 2 Arsenic concentration ratio in P. vittata pinnae
2. 2 AmountofAsaccumulationinpinnae
There were differences among the pinnae in terms
of the amount of As accumulation，which was based on
As concentration and biomass dry weight （Fig .3 ） .
For brevity，the data for biomass dry weight of pinnae
is not shown. The 10 sampled pinnae could be divided
into two groups based on their As accumulation： low-
As accumulation （mean value<5 g），and high-As ac-
cumulation （mean value>8 g）. The As accumulation
was greater in middle pinnae and lower in the basal
and top pinnae. In addition，there were large differ-
ences in As enrichment between low-As P. vittata and
high-As P. vittata. The amount of As accumulation in
the low -As P. vittata gradually increased along the
frond and then decreased slightly. The dynamic equa-
tion of heavy metals parabolic diffusion equation was
A
rs
en
ic
co
nc
en
tra
tio
n
ra
tio
n
Pinnae order of low-arsenic P. vittata（a）
Y=1.05-0.05X+0.003X2
R2=0.98
Pinnae order of high-arsenic P. vittata（b）
A
rs
en
ic
co
nc
en
tra
tio
n
ra
tio
n
Y=1.05-0.05X+0.002X2
R2=0.99
757· ·
南 方 农 业 学 报 46卷
used to simulate the data of As accumulation. The re-
sults showed that the fitting effect was good，the corre-
lation coefficient R2 in the dynamic equation of low-
As fronds was 0.88，fitting the equation： Y=0.48 +
0.098X-0.003X2.
However，in the high-As P. vittata，the amount of
As accumulation increased along the frond and then
decreased significantly in the top pinnae. Based on
biomass and As concentration，As accumulation in the
middle pinnae was 3.1 times and 2.8 times higher than
in the basal and top pinnae，and the correlation coeffi-
cient R2 in the dynamic equation of high -As fronds
was 0.89，fitting the equation： Y=0.67+0.10X-0.005X2.
Therefore，from what has been showed above，there was
a “∩”-type distribution of As accumulation along the
frond. These trends in distribution showed that there
was a high accumulation of As in the middle pinnae.
3 Discussion
The results of this study indicated that both the
low As and high As P. vittata，As concentration in the
pinnae showed that it was high in the basal pinnae，
with the increase of leaf position，As concentration re-
duced gradually，and then increased in the top pinnae.
The results verified that previous research had report-
ed. The As concentration increased from 3000 mg/kg
in the top pinnae to 6000 -9000 mg/kg in the basal
pinnae （Lombi et al.，2002），the variation was up to 3
times. The differences of As concentration of the top
pinnae and the basal pinnae would be about 1.5 times
in the different pinnae of the same frond. The distri-
bution pattern suggested that As was transported and
unloaded from the basal pinnae to the middle pinnae，
possibly reflecting the diffusion of As from a region of
high concentration to one of lower concentration. The
fact that the concentration of As was higher in the top
pinnae than that of the middle pinnae may reflect the
strong physiological activities of the new pinnae at the
top of the frond，which would produce a strong tran-
spiration pull and/or have strong metabolic activity
with more energy and greater enzyme activity.
This rule also suggested that As Xylem transport
capacity along the vascular bundle to the blade tip
was very strong. From the point of whole plant，As also
have strong ability of transportation to the ground （Xi-
ao et al.，2006）. As transport characteristics of P. vit-
tata indicated that it was an efficient process for As
transportation in the xylem. It was generally believed
（Yang et al.，2002） that the cation exchange capacity
in the xylem cell walls was high，which block the met-
al cation transport upwards. As existed as arsenous
acid radical anions form in the P. vittata （Huang et
al.，2004b），which led to the block effect for As was
weak in the xylem，and most As could transport to the
above ground part in the xylem. This may be one of
the important mechanisms that As accumulation in P.
vittata. At the same time other studies showed that As
in the midrib with strong ability to transport to the
mesophyll tissue，which mean that As was with strong
ability to unload in the xylem （Chen et al.，2004）.
This study found that the concentration of As diluted
in middle pinnae objectively due to its larger biomass.
So As concentration in the middle pinnae was lower
than that in basal or top pinnae.
It was very important to evaluate the phytoreme-
diation efficiency in phytoremediation projects，and
how to collect the samples in order to achieve scientif-
ic and accurate evaluation of the phytoremediation ef-
ficiency in phytoremediation projects was also impor-
tant （Chen et al.，2007）. It was generally believed that
collecting the middle pinnae was appropriate. But the
study found that although As accumulation amount the
middle pinnae was larger，As concentration was rela-
Fig.3 Arsenic enrichment amount ratio in pinnae of P. vittata
Y=0.48+0.098X-0.003X2
R2=0.88
A
rs
en
ic
en
ric
hm
en
ta
m
ou
nt
ra
tio
Pinnaeorderoflow-arsenicP.vitata（a） Pinnaeorderofhigh-arsenicP.vitata（b）
Y=0.67+0.10X- 0 5X2
R2=0.88
A
rs
en
ic
en
ri
ch
m
en
ta
m
ou
nt
ra
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Journal of Southern Agriculture758· ·
5期
tively low，So only collecting the middle pinnae could
not reflect the phytoremediation efficiency. Samples
were collected to evaluate phytoremediation efficiency
accurately，the middle pinnae，the basal pinnae and
top pinnae should be collected. Meanwhile，to improve
the phytoremediation efficiency，measures should be
taken to improve P. vittata biomass（Liao et al.，2007），
specially improve the middle pinnae biomass.
4 Conclusions
The result shows that to evaluate phytoremedia-
tion efficiency accurately，the middle pinnae in com-
pany with the basal pinnae and top pinnae should be
collected. Meanwhile，to improve the phytoremediation
efficiency，measures should be taken to improve P.
vittata biomass，specially improve the middle pinnae
biomass. The results of these studies will help to de-
sign appropriate strategies for collection of plant sam-
ples in phytoremediation projects，allowing accurate e-
valuation of phytoremediation efficiency. In addition，
these results will be useful for determining the appro-
priate planting density of hyperaccumulator.
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（责任编辑 汪羽宁）
WANG et al.：Accumulation of arsenic and its distribution in pinnae of hyperaccumulator Pteris vittata L. 759· ·