INTERNATIONAL JOURNAL OF BUSINESS, SOCIAL AND SCIENTIFIC RESEARCH

ISSN: 2309-7892, Page: 87-94, Volume 01, Issue 02, January-March 2014
Review Paper
MITIGATION OF SOIL ARSENIC BY PHYTOREMEDIATION PROCESS

U. Mayda1, M.A. Akond1, M.I. Manirul2, H. Mehraj3 and AFM Jamal Uddin3*

U. Mayda, M.A. Akond, M.I. Manirul, H. Mehraj and AFM Jamal Uddin (2014). Mitigation of Soil Arsenic by
Phytoremediation Process. Int. J. Bus., Soc. and Sci. Res. 01(02):87-94. Retrieve from
http://www.ijbssr.com/currentissueview/13090115

Received Date: 22/01/2014 Acceptance Date: 31/01/2014 Published Date: 07/02/2014

Abstract
An experiment was conducted at the Department of Botany, Jahangirnagar University,
Dhaka, Bangladesh to mitigate arsenic from soil by using phytoremediation process
during the period from October 2012 to April 2013 following Completely Randomized
Design (CRD) with three replications. Four non edible fern (viz. V1, Adiantum sp; V2,
Microlepia sp; V3, Pteris vittata and V4, Christella sp) and four soil arsenic
concentrations (viz. A0, Control; A1, 1000-ppm; A2, 2000-ppm and A3, 4000-ppm) were
exploited. Pteris vittata L. was found as the arsenic hyperaccumulating plant. It can
survive with a very high concentration of arsenic in soil and accumulate 27829.7-ppm,
23274.7-ppm and 14911.0ppm arsenic when soil was treated with 4000-ppm, 2000-ppm
and 1000-ppm arsenic respectively. On the other hand other fern species used in the
experiment could not survive in soil treated with arsenic. Throughout the experimental
period, different growth parameters of fern species showed distinct variation with
increasing concentration of arsenic. Phytoremediation of arsenic by using Pteris vittata
is a possible way for the mitigation of arsenic form soil which can keep away of arsenic
pollution in food chain.

Key words: Pteris vittata, soil arsenic and phytoremediation.

Introduction
Arsenic (As), a metalloid element is known as poison and its contamination is one of the
promising public concerns not only in Bangladesh but also in whole world due to its toxic and
carcinogenic nature. Arsenic in soil and groundwater can be taken up by plants that are directly or
indirectly ingested by animals and humans. Widespread use of groundwater for irrigation is a
route of arsenic contamination into food chain (Huq et al. 2006) and presence of arsenic in food
chain (Duxburya et al. 2003, Das et al. 2004) affect human health (Zhao et al. 2009) directly or
indirectly. High level of arsenic in irrigated water and soil appears to result in higher
concentration of arsenic in root, stem and leaf of rice plants (Abedin et al. 2002, Das et al. 2004)
and dietary intake of rice is potentially a major As exposure pathway in countries (Smith et al.
2008) like Bangladesh. Arsenic in irrigation water is responsible for arsenic accumulation (Azad
et al. 2013) into plants. Arsenic contamination in soil increases day by day. It is important to
remove arsenic from top soil and prevent possible impacts on the ecosystems. Removal of arsenic
from soil by using phytoremediation and render to enter into food chain. Many non edible fern
species like Pteris vittata, Adiantum sp., Christella sp., Microlepia sp. etc have great potentiality
in arsenic phytoremediation and also available in Bangladesh. to the lack of surface water,
insufficient rainfall and soluble form of arsenic increase its available forms in soil; are uptaken by
plant root with other nutrient. Red amaranth uptake 41.8g/gm (leaf), 69.7g/gm (stem), and
8.7g/gm (root) when it grows on 15mg/L arsenic (Choudhury et al., 2009), also many report
suggest that many edible plants uptake huge amount of arsenic that means entrance of arsenic into
the vegetables, fruits, cereals etc. These foods are sold in Agora, Nandon, Minabazar, Shawpno,
so many other super shop and local food market for human consumption, are harmful for human
health. Keeping this point in view, using non edible plant material which can remove arsenic from
soil and can stop to the entrance of arsenic into the food chain and providing a low cost tool for
the arsenic removal to the farmers level.

*Corresponding Author: jamal4@yahoo.com

1
Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh;
2
Department of Agricultural Chemistry, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh.
3
Department of Horticulture, Sher-e-Bangla Agricultural University, Dhaka, Bangladesh.
Mayda et al. 88

Materials and Method

The experiment was conducted at the Jahangirnagar University (JU), Savar, Dhaka, Bangladesh
during October 2012 to April 2013 with Complete Randomized Design (CRD) using three
replication. Four non-edible fern species viz. V1: Adiantum sp., V2: Microlepia sp., V3: Pteris
vittata and V4: Christella sp and four arsenic concentrations viz. C0: Control, A1: 1000-ppm, A2:
2000-ppm and A3: 4000-ppm was exploited on experiment. Four non-edible fern species (Plate 1)
were collected from different sites of the JU campus. Approximately 3 weeks old plantlets were
transplanted into the experimental pots. Pots were prepared 15 days before transplantation.
Weeding and stubbles removals were performed manually before transplanted such 7 days
interval after transplantation. Arsenic was applied in the form of Arsenic trioxide (As2O3) which
was purchased from Loba Chemie Pvt. Ltd., India. As per analysis of molecular weight 1.32 g of
As2O3 contains 1 gm As hence following amounts of As2O3 viz. 0 g, 5.28 g, 10.56 g, and 21.12 g
for control, 1000-ppm, 2000-ppm and 4000-ppm respectively for the treatments of arsenic in the
fern As2O3 applied into pot and then mix with soil by using stick. Data was taken on following
parameters plant height, leaf numbers, leaf length, leaflet numbers, leaf area, plant leaf biomass,
arsenic accumulation (0.5 mg tested sample), plant survival period. Leaf area was measured by
non-destructive method using CL-202 Leaf Area Meter (USA). After growing fern species all fern
leaves are collected; dried and smashed by mortar and pastel machine for chemical analysis in
Bangladesh Council of Scientific Research Institute (BCSIR), Dhaka, Bangladesh. Chemical
analysis of arsenic used Atomic Absorption Spectrometer where use argon for carrier gas and
arsenic was melted by 925C; was approved by ISO organization.
50 times dilution: 5 ml concentrated HCl was taken at 50 ml volumetric flasks for transferring
arsenic into arsenic trioxide and a little bit of distilled water was added. Then 1ml solution was
taken very carefully from each volumetric flask to avoid bubble and KI (1 gm) wash added in
solution with 150 ml distilled water. After that 0.5 gm sample was taken in volumetric flask and
mixed distilled water up to 50 ml and solution turned into yellow color. Another volumetric flask
made blank solution, where contain only HCl, KI and distilled water for arsenic analysis.
1000 time dilution: 2 ml solution was taken into 500 volumetric flasks, mixed with distilled water
up to 500 ml and shaken very carefully. Then 5 ml solution was taken into 250 ml volumetric
flask and mixed with distilled water up to 250 ml. Again, 5 ml solution from 250 ml solution was
taken into 25 ml volumetric flask then 2.5 ml HCl and 2.5 ml KI was added and mixed distilled
water, shaking was done very smoothly until turn it into yellow colour. Standard solution arsenic
was added with HCl 2, 5, 10, 15, 20 ppb respectively.
5000 time dilution: Similar to the 1000 time dilution. But 4 ml solution was taken in case of 2ml.
5 ml HCl and KI was added into 5000 ml volumetric flask, solution was made into 25 ml and 5 ml
was taken in flask.
Collected data were statistically analyzed using MSTAT-C computer package programme. Mean
for every treatments were calculated and the analysis of variance for each one of the characters
was performed by Ftest (Variance Ratio). Difference between treatments was evaluated by LSD
test at 5% level of significance (Gomez and Gomez, 1984).

(a) (b)

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Mitigation of Soil Arsenic 89

(c) (d)
Plate 1. Fern species a) V1; Adiantum sp., b) V2; Microlepia sp. c) V3; Pteris
vittata, d) V4; Christella sp.
Result and Discussion
Significant variation was found among the fern species performance in terms of leaf number. Leaf
number showed statistically significant differences among V1, V2, V3 and V4 at 7, 14, 21 and 28
DAT (Fig. 1). Highest number of leaf (10.9) was obtained from V3 (Pteris vittata) and lowest
number of leaf (3.2) was obtained from V4 (Christella sp.) fern at 28 DAT. Leaf number of fern
species showed statistically significant differences among control, 1000-ppm, 2000-ppm, 4000-
ppm at 7, 14, 21 and 28 DAT (Fig. 2). Highest leaf number (14.2) was observed in control treated
plants and lowest leaf number was found in 4000-ppm (1.9) at 28 DAT. Leaf number of fern
species showed statistically significant differences among treatment combinations at 7, 14, 21 and
28 DAT (Fig. 3). Highest number of leaf (15.7) was found under the V3A0 treatment and lowest
number of leaf (0.0) was found under V2A0, V4A3 and V1A3 treatment at 28 DAT.
Significant variation was found among the fern in respect of leaf area (cm2). Leaf area of fern
species showed statistically significant differences among V1, V2, V3 and V4 at 0, 7, 14 and 21
DAT (Fig. 4). Highest leaf area (19.3 cm2) was obtained from V3 (Pteris vittata) and lowest (12.8
cm2) was obtained from V4 (Christella sp.) fern at 21 DAT. Leaf area (cm2) was significantly
affected by arsenic treatments. Leaf area of fern species showed statistically significant
differences among control, 1000-ppm, 2000-ppm and 4000-ppm at 0, 7, 14 and 21 DAT (Fig. 5).
Highest leaf area (18.1 cm2) was observed in control treated plants and lowest (12.9 cm2) leaf area
was found in 4000-ppm at 21 DAT. Leaf area showed statistically significant differences among
treatment combinations at 0, 7, 14, and 21 DAT (Fig. 6). Highest leaf area (22 cm2) was found
under the V3A0 treatment and lowest (10.7 cm2) found under V4A3 treatment at 21 DAT.

12.0 16.0
A0
V1
A1
V2
12.0 A2
V3
Number of leaves.

Number of leaves.

8.0 A3
V4
8.0

4.0
4.0

0.0 0.0
7 14 21 28 7 14 21 28
Days after transplanting (DAT) Days after transplanting (DAT)

Fig.1. Effect of fern species on number of leaves Fig. 2. Effect of arsenic concentrations on
at different days after transplanting (DAT) number of leaves at different days after
transplanting (DAT)

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Mayda et al. 90

18.0
7 DAT 14 DAT 21 DAT 28 DAT

12.0
Number of leaves.

6.0

0.0
V1A0 V1A1 V1A2 V1A3 V2A0 V2A1 V2A2 V2A3 V3A0 V3A1 V3A2 V3A3 V4A0 V4A1 V4A2 V4A3

Combination

Fig. 3. Combined effect of fern species and arsenic concentrations on number of leaves at different
days after transplanting (DAT)

V1, Adiantum sp; V2, Microlepia sp; V3, Pteris vittata; V4, Christella sp; A0,Control; A1,1000ppm;
A2, 2000ppm and A3, 4000ppm

20.0 20.0
V1 A0
V2 A1
V3 A2
V4 A3
15.0
L eaf area (cm 2 )

15.0
Leaf area (cm2)

10.0 10.0

5.0 5.0
0 7 14 21 0 7 14 21
Days after transplanting (DAT) Days after transplanting (DAT)

Fig.4. Effect of fern species on leaf area at Fig. 5. Effect of arsenic concentrations on leaf
different days after transplanting area at different days after transplanting
(DAT) (DAT)

25.0
0 DAT 7 DAT 14 DAT 21 DAT

20.0
Leaf area (cm2)

15.0

10.0

5.0
V1A0 V1A1 V1A2 V1A3 V2A0 V2A1 V2A2 V2A3 V3A0 V3A1 V3A2 V3A3 V4A0 V4A1 V4A2 V4A3

Combination

Fig. 6. Combined effect of fern species and arsenic concentrations on leaf area at different days
after transplanting (DAT)

V1, Adiantum sp; V2, Microlepia sp; V3, Pteris vittata; V4, Christella sp; A0,Control;
A1,1000ppm; A2, 2000ppm and A3, 4000ppm

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Mitigation of Soil Arsenic 91

Plant leaf biomass varied significantly among the fern species. Among the fern species V3 (Pteris
vittata) provides maximum plant biomass (1601.7 mg) followed by V2 (1001.7 mg) whereas
minimum from V1 (439.2 mg) (Table 1). Reduction of plant leaf biomass denotes that low plant
growth. Here, Pteris vittata provided maximum plant leaf biomass i.e., Pteris can grow well in
arsenic contaminated soil. Different arsenic concentration showed significant variation in terms of
plant leaf biomass. Maximum plant leaf biomass (1339.2 mg) was recorded from control (A0;
0ppm) whereas the minimum plant leaf biomass (534.2 mg) was found from 4000-ppm (A3)
arsenic treatment (Table 2). The present study identified that plant leaf biomass was reduced with
the increase of arsenic concentration. Plant biomass varied due the variation of the concentration
of arsenic (Wei et al., 2006). Combined effect also showed significant difference for plant leaf
biomass. Maximum plant biomass (2301.7 mg) was found from V3A0 and the minimum plant
biomass (201.7 mg) was recorded from V1A3 (Table 3).
Arsenic accumulation by fern species was varied significantly. From the each treatment, 0.5 mg
of sample was tested in each case and highest arsenic accumulation (28652.3-ppb) was found
from V3 (Pteris vittata) whereas minimum from V1 (Adiantum sp) (14425.3-ppb) (Table 1). This
result shows that Pteris vittata has potentiality to uptake maximum amount of arsenic from
arsenic contaminated soil. The ability of P. vittata to translocate arsenic from the roots to the
fronds (7377% arsenic in the fronds), reduce arsenate to arsenite in the fronds (>50% AsIII in
the fronds) (Luongo and Ma, 2005). Different concentration significantly influenced the arsenic
accumulation from contaminated soil. 4000-ppm treated plants uptake maximum amount of
arsenic (45739.8-ppb) while minimum amount of arsenic accumulation (2912.2-ppb) was
obtained from the control treatment (Table 2). This result indicated that ferns grown with 4000-
ppm arsenic treated soil uptake maximum amount of arsenic. Combined effect showed
statistically significant variation in arsenic accumulation. Maximum accumulation (55652.3-ppb)
was recorded from V3A3 and minimum from V2A0 (454.3-ppb) (Table 3).
Table 1. Responses of fern species against arsenic to different attributes

Arsenic accumulation on Total arsenic

Plant leaf biomass
Variety 0.5 mg tested sample accumulation on
(mg)
(ppb) plant leaves (ppm)
V1 439.2 d 14425.3 d 1960.1 d
V2 1001.7 b 16890.3 c 5326.3 c
V3 1601.7 a 28652.3 a 18398.9 a
V4 646.7 c 21327.3 b 5583.0 b
LSD0.05 1.7 2.1 4.7
CV% 0.2 0.1 0.1
Table 2. Fern responses of arsenic concentrations to different attributes

Arsenic accumulation on Total arsenic

Plant leaf biomass
Treatments 0.5 mg tested sample accumulation on
(mg)
(ppb) plant leaves (ppm)
A0 1339.2 a 2912.2 d 2473.9 d
A1 1064.2 b 9136.7 c 5689.6 c
A2 751.7 c 23506.5 b 10060.2 b
A3 534.2 d 45739.8 a 13044.7 a
LSD0.05 1.7 2.1 4.7
CV% 0.2 0.1 0.1

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Mayda et al. 92

Table 3. Combined responses of fern species and arsenic concentration to different attributes

Arsenic accumulation on Total arsenic

Plant leaf biomass
Combination 0.5 mg tested sample accumulation on
(mg)
(ppb) plant leaves (ppm)
V1A0 651.7 j 864.8 o 285.0 p
V1A1 501.7 k 2614.8 n 657.8 n
V1A2 401.7 m 14669.0 i 2938.0 k
V1A3 201.7 n 39552.3 d 3959.7 j
V2A0 1501.7 c 454.3 p 343.7 o
V2A1 1201.7 e 2977.3 m 1789.7 l
V2A2 801.7 h 20877.3 g 8354.7 f
V2A3 501.7 k 43252.3 c 10817.2 d
V3A0 2301.7 a 6589.8 k 7580.3 g
V3A1 1801.7 b 16564.8 h 14911.0 c
V3A2 1301.7 d 35802.3 e 23274.7 b
V3A3 1001.7 f 55652.3 a 27829.7 A
V4A0 901.7 g 3739.8 l 1686.6 M
V4A1 751.7 i 14389.8 j 5400.0 I
V4A2 501.7 k 22677.3 f 5673.5 H
V4A3 431.7 l 44502.3 b 9572.2 E
LSD0.05 3.5 4.2 9.5
CV% 0.2 0.1 0.1

V1, Adiantum sp; V2, Microlepia sp; V3, Pteris vittata; V4, Christella sp; A0,Control;
A1,1000ppm; A2, 2000ppm and A3, 4000ppm

Table 4. Plant survival period at different treatment combinations

Treatment Combination Plant Survival Period
V1A0 Stay Alive
V1A1 35 Days
V1A2 30 Days
V1A3 27 Days
V2A0 Stay Alive
V2A1 33 Days
V2A2 29 Days
V2A3 26 Days
V3A0 Stay Alive
V3A1 Stay Alive
V3A2 Stay Alive
V3A3 Stay Alive
V4A0 Stay Alive
V4A1 35 Days
V4A2 29 Days
V4A3 25 Days

V1, Adiantum sp; V2, Microlepia sp; V3, Pteris vittata; V4, Christella sp; A0,Control;
A1,1000ppm; A2, 2000ppm and A3, 4000ppm

This research work exhibited distinct variations in total arsenic accumulation of fern
species. Maximum accumulation (18398.9-ppm) was found from in V3 (Pteris vittata) and
minimum (1960.1-ppm) were obtained from V1 (Adiantum sp) (Table 1). Gonzaga et al. (2008)
also reported that Pteris vittata can be used to remediate arsenic-contaminated soils by repeatedly
harvesting its fronds. Total arsenic accumulation on plant leaves varied significantly among the
soil treated with different arsenic concentrations. Maximum accumulation (13044.7-ppm) was
found from soil treated with 4000-ppm arsenic whereas minimum from A0 (2473.9-ppm) (Table
2). Soil treated with 4000-ppm arsenic hastened arsenic uptake. Combined effect influenced total
arsenic accumulation on leaves of ferns. Maximum arsenic uptake was found from V3A3
(27829.7-ppm) followed by V3A2 (23274.7-ppm) and V3A1 (14911.0ppm) and minimum from
V1A0 (285.0-ppm) (Table 3). Wang et al. (2002) also found that Pteris vittata accumulated
arsenic in the fronds up to 27,000 mg arsenic kg-1(ppm) dry weight.
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Mitigation of Soil Arsenic 93

From the study it was observed that all the ferns in the soil without arsenic treatments
i.e., control were alive (Plate 2). But ferns in the soil treating with arsenic at different
concentrations were dead except Pteris vittata. It was found that Pteris vittata were survive after
the soil treated with very high concentrations of arsenic (Table 4). So it can be used as
hyperaccumulators for arsenic.

(a) (b)

(c) (d)

Plate 2. Effect of arsenic concentration at 36 days after transplanting for different fern species
at 1000ppm, 2000ppm and 4000ppm

(a) V1; Adiantum sp., (b) V2; Microlepia sp. (c) V3; Pteris vittata, (d) V4; Christella sp.

Conclusion
From above results it was observed that Pteris vittata (V3) fern bears maximum arsenic
uptake ability from soil and also varied due to arsenic concentration in soil. Considering above
circumstances, it can be concluded that Pteris vittata (V3) is the most excellent fern for arsenic
uptake from soil and it can alive when soil arsenic concentration is 4000ppm. In our country,
highly arsenic affected soil also area where soil arsenic is gradually increased can be reduced by
Pteris vittata. So, result of present study offers a good scope for mitigation from soil by using this
methods and good result found in Pteris vittata. But further research could be conducted and after
consecutive trial it is possible to protect the entrance of arsenic into food chain.
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