CSIRO PUBLISHING

Australian Journal of Botany

Communication
http://dx.doi.org/10.1071/BT15066

Chromium and nickel accumulation in the macrophytes

of the Kawasi wetland on Obi Island, North Maluku
Province, Indonesia

R. Amin A,C, M. Edraki A, D. R. Mulligan A and T. H. Gultom B

A
Centre for Mined Land Rehabilitation, Sustainable Minerals Institute, The University of Queensland,
Brisbane, QLD 4072, Australia.
B
Harita Nickel Group, Ratu Plaza Office Tower, 14th floor, Jl. Jend. Sudirman kav. 9, Jakarta Pusat, Indonesia 10270.
C
Corresponding author. Email: r.amin@uq.edu.au

This paper originated from the special issue ‘Ultramafic Ecosystems: Proceedings of the 8th
International Conference on Serpentine Ecology (Part 2)’.

Abstract. Five macrophytes, namely Crinum asiaticum L. (Amaryllidaceae), Lepironia articulata (Retz.) Domin
(Cyperaceae), Machaerina rubiginosa (Spreng.) T. Koyama (Cyperaceae), Pandanus sp. (Pandanaceae) and Nepenthes
mirabilis (Lour.) Druce (Nepenthaceae), were identified in the Kawasi wetland, a natural wetland on Obi Island, Indonesia,
that overlies ultramafic rocks. The dominant species in this wetland was C. asiaticum, a native of the region. The surface
runoff in the catchment of the Kawasi wetland was derived from serpentine soils, areas of which were being mined for
nickel and, as a result, the water that flowed to the wetland typically contained dissolved chromium and nickel. In this study
we investigated the accumulation of chromium and nickel in the macrophytes of the wetland. The five species of macrophytes
under investigation accumulated greater quantities of chromium and nickel in their roots than in their shoots, with Pandanus
sp. having the highest translocation factor (as evidenced by the highest shoot : root ratio) for both chromium and nickel.
The species with the highest concentrations of the metals in both roots and shoots was C. asiaticum.

Additional keywords: chromium, Nepenthes mirabilis, wetlands, ultramafic.

Received 17 October 2014, accepted 25 July 2015, published online 14 September 2015

Introduction some level of trivalent chromium for metabolism and growth

Nickel laterite mines in Indonesia must undertake rigorous
environmental protection and management measures because
the potential risks of their operations to the environment are
high. The nickel-enriched laterite weathering profiles derived
from serpentine rocks are a focal point for the laterite nickel
miners, but nickel-laterites can also contain chromium, among
other metals (Ahmad 2009). When the subsurface horizons are
exposed to mining, chromium can be released or leached into the
environment. In nature, chromium usually exists in two states:
hexavalent and trivalent (Tu et al. 2013). Chromium can also
occur in divalent and pentavalent forms (Michailides et al. 2013).
Between the hexavalent and trivalent states, the form of primary
environmental concern is hexavalent chromium [Cr(VI)], which
can be extremely toxic to plants, animals and humans (Blowes
2002; Ahmad 2009). The toxicity of hexavalent chromium is one
hundred-fold higher than the toxicity of trivalent chromium
(Michailides et al. 2013). It is carcinogenic (Blowes 2002) and
has been causally linked to liver and bone damage/disease in
humans, and the blocking of functional groups of vital enzymes
(Moore 1991). However, both plants and animals may require
(Tirez et al. 2007).
Wetlands have proven to be effective in removing or reducing
a range of pollutants that can often be found in excess in
wastewaters from a range of contaminant sources (Sheoran and
Sheoran 2006). These systems are seen as a preferred treatment
option over more complex engineering solutions because they are
more environmentally friendly (and are an attractant to both birds
and other fauna), provide aesthetic value to an area, can be at least as
effective in contaminant management, and are comparatively less
expensive to both construct and maintain (Michailides et al. 2013).
Several studies in metal-polluted industrial areas have proven
wetlands to be successful in removing pollutants, and these
include the Swabi district in Pakistan (Khan et al. 2009), the
Kanpur district in India (Gupta et al. 2011) and the Tennessee
Valley in the USA (Mays and Edwards 2001). However, reported
studies on wetland performance in Indonesia, and certainly in
North Maluku Province, are scarce.
In a wetland system, a combination of biological, chemical,
and physical processes are involved in ameliorating the elevated
levels of metals that can be present in drainage from contaminated

Journal compilation
CSIRO 2015 www.publish.csiro.au/journals/ajb

B Australian Journal of Botany R. Amin et al.

sites. One of the key biological processes is metal accumulation the species with the highest levels of metal accumulation (because
in plant tissues (Pourkhabbaz et al. 2011; Yadav and Chandra the IVI does not measure elemental concentrations).
2011), and in the present study we focused on the roles and The locations of sampling plots were determined based on
contributions of wetland macrophytes in this process. accessibility and safety considerations. The water and sediment
The use of native macrophytes was an important consideration depth meant sampling could only be conducted from a boat, and
in this study due to the ecosystem damage that introduced the density of vegetation further restricted access to, and hence,
macrophytes can bring. Invasion of the study area by introduced sampling of, the entire area of the wetland.
species has occurred, and species such as Eichhornia crassipes The sampling plot area chosen was 1
1 m, which is similar or
(Mart.) Solms. (water hyacinth), Hydrilla verticillata (L.f.) Royle larger than the size range used by Carvalho et al. (2013), Vymazal
(hydrilla) and Lythrum salicaria L. (purple loosestrife) (Cronk et al. (2009) and Di Luca et al. (2011). Given the nature of the
and Fennessy 2001) are common, as they occur across wetlands environment and vegetation assemblages, which was visually
throughout Indonesia. In selecting the native species for this homogenous (Fig. 1), it was considered that larger plots would
study, those that could easily grow and spread in unfavourable be more suitable for this current study.
environmental conditions were further criteria for selection In the on-site laboratory, the plant samples were washed using
(Srivastav et al. 1994). running tap water to remove sediment and mud, especially from
the roots. After washing, the samples were rinsed with deionised
Materials and methods (DI) water and blotted dry using tissue paper. Each plant sample
Wetland description was divided into two; one for preservation and pressing for
species identification and deposit in the herbarium, and the
The Kawasi wetland is a freshwater body located 200 m from the
other for chemical analysis. The plant samples destined for the
western coast of Obi Island, North Maluku Province, Indonesia.
herbarium had complete and undamaged plant parts and were
It is approximately 100 ha in area, expanding in width at the
preserved by spraying with 70% alcohol. The plant samples
northern end. The wetland has two principal influents; the
destined for chemical analysis were kept at low temperature
Toguraci River, and the runoff from a causeway that was
during storage and transfer to an external independent laboratory
constructed across the wetland to connect the mainland and a
in Manado, North Sulawesi Province.
port, which was operated and managed by the company. The
effluent of the wetland flows back into the Toguraci River
downstream and then to the ocean. The wetland overlies Methods for analysing the samples
ultramafic rock that is rich in nickel, and several nickel mining
The plant tissue samples were analysed in WLN Indonesia, which
operations are in the vicinity of the wetland. The mining activities
was an independent laboratory situated in Manado, Indonesia,
commenced in early 2000 and exposure of serpentine soil and the
using ICP-MS (Thermo
Series II-Thermo German, Thermo
consequent leaching and runoff have resulted in a potential increase
Fisher Scientific, Bremen, Germany) for metals. Samples were
in the levels of nickel and chromium, in particular, flowing into the
rinsed using DI water, dried using absorbent paper, and separated
wetland.
into roots and shoots. The digestion of the fresh samples was
conducted in three stages. In the first stage, the samples were
Sample preparation
digested using concentrated HNO3 at room temperature for 6 h.
The study was conducted during the rainy season of August– The second stage consisted of heating the samples at 85C for 2 h.
September 2013. Ten sampling plots, which were 1
1 m2, from While cooling, the third stage of the digestion was initiated and
locations across the wetland were selected and, in each plot, plant 30% H2O2 was added to the samples followed by a further heating
species were identified, numbers of plants were recorded, and
plant samples for tissue analysis and species identification were
collected. If there were two species of macrophyte in one plot,
samples of both were collected. The LIPI-Herbarium Bogoriense
Indonesia conducted the species identification from the
herbarium specimens that were prepared on site. Other data
from the sampling plots, including the coverage of species (by
visual estimation) and the number of plots containing a given
species, were used to determine the importance value index (IVI)
(Jafari Ghavzan et al. 2006):
The importance value index of macrophytes was calculated:
IVI ¼ relative frequency ðRFÞ þ relative coverage ðRCÞ; ð1Þ

where RF is the number of plots in which a given species was

encountered divided by the total number of plots, and RC is the
total coverage of species divided by the sum of all coverage of all
species. The calculation of IVI was to measure the dominance of
a given species in the wetland. The IVI value may not be directly
related to the response of plants to the chemistry of the water
body, meaning that the most dominant species is not necessarily Fig. 1. Kawasi wetland is dominated by Crinum asiaticum.
Chromium and nickel accumulation in macrophytes, Indonesia Australian Journal of Botany C

step of 85C for 1 h. To obtain a clear solution, the samples were
filtered and diluted to 50 mL with DI water.
Results
Plant species of Kawasi wetland
Visually, only one species dominated the Kawasi wetland area,
namely C. asiaticum (Fig. 1), although there were typically
up to two to three different species identified in each sampling
plot. In total, there were five macrophytes species found in the
wetland: C. asiaticum, L. articulata, M. rubiginosa, Pandanus sp.
and N. mirabilis. The phenotype of C. asiaticum present had
noticeably broad leaves whereas M. rubiginosa had narrow leaves
that were not that dissimilar from the stems of L. articulata.
Pandanus sp. and N. mirabilis were the least dominant species in
this wetland, but these species were regularly found in the less
permanently inundated areas on the Island, namely the flood
plains and riverbanks.
Most macrophytes in these environments regenerate from
rhizomes growing underwater and in this study, C. asiaticum,
L. articulata and M. rubiginosa all had extensive roots and
rhizomes underwater. In comparison, the roots of Pandanus
sp. and N. mirabilis were generally only partially submerged.
Personal communications with the local people of Obi Island
led the researchers to conclude that the C. asiaticum in this
wetland, which has white flowers, never bloomed. However,
there was little information about the flowering period for other
species provided by local people. At the time of sampling for this
study, no flowers or fruits were encountered.
Based on the vegetation metrics and analyses that were
undertaken, Table 1 shows the importance value index (IVI) of
each species.
The IVI was calculated by combining two factors – relative
frequency and relative cover (Eqn 1). This calculation results in a
total IVI value of 200%. Macrophyte density per se was not
measured in this survey as it would be a misleading number given
that regeneration of the macrophytes occurs from underwater
rhizomes and hence the ability to identify individual plants is
problematic.
Concentration of metals in plant tissues
In total, there were 22 plant samples collected from 10 sampling
plots in the wetland, consisting of C. asiaticum (n = 10), and
M. rubiginosa and L. articulata (both n = 5). However, there was
only one sample of Pandanus sp. and N. mirabilis encountered
across all the plots. Given the high bioavailability of nickel and
chromium in serpentine soils (Baker 1981), Table 2 shows the
concentrations of these elements in the roots and shoots of the
macrophyte species sampled from the Kawasi wetland.
Tappero et al. (2007) reported that for species to be considered
in the category of hyperaccumulators, plants would likely have
a translocation factor (TF) >1. The TF is the ratio between
concentration (g kg–1 DW) of a given element in shoot and in
roots. The results are shown in Table 3.
The highest TF of 0.52 was recorded for the Pandanus sp., and
this applied to both nickel and chromium. However, this finding is
of limited value since it was derived from a single sample. The

Table 1. Importance value index of the wetland plant species

Abbreviations: RF, relative frequency; RC, relative coverage; IVI, importance value index

Species Family Frequency RF Coverage RC IVI

Crinum asiaticum Amaryllidaceae 1.00 45.5% 160 91.2% 136.6%
Lepironia articulata Cyperaceae 0.60 27.3% 3.0 1.7% 29.8%
Machaerina rubiginosa Cyperaceae 0.40 18.2% 4.5 2.5% 19.9%
Pandanus sp. Pandanaceae 0.10 4.5% 4.0 2.3% 6.8%
Nepenthes mirabilis Nepenthaceae 0.10 4.5% 4.0 2.3% 6.8%
Total 2.20 175.5 200.0%

Table 2. The mean concentration of chromium and nickel in macrophytes (mg kg–1 DW þ s.d.)

Lepironia articulata Crinum asiaticum Machaerina rubiginosa Pandanus sp. Nepenthes mirabilis
Average in roots
Chromium 55.5 ± 30.4 130.5 ± 151.5 124.3 ± 146.7 37.1 180.8
Nickel 366.5 ± 191.5 942.8 ± 1143.0 1115.3 ± 1292.1 226.6 678.4
Average in shoots
Chromium 10.5 ± 12.5 23.7 ± 13.0 10.9 ± 3.9 19.4 14.3
Nickel 38.6 ± 51.1 140.3 ± 104.3 45.1 ± 33.3 118.1 36.9

Table 3. Translocation factor (TF) of macrophytes in the Kawasi wetland

TF Lepironia articulata Crinum asiaticum Machaerina rubiginosa Pandanus sp. Nepenthes mirabilis
Chromium 0.19 ± 0.41 0.18 ± 0.10 0.09 ± 0.03 0.52 0.08
Nickel 0.11 + 0.27 0.15 ± 0.09 0.04 ± 0.03 0.52 0.05
D Australian Journal of Botany
Fig. 2. Roots and rhizomes of Lepironia articulata found underwater in the
wetland.
lowest TF for nickel was in M. rubiginosa, and for chromium
was in N. mirabilis, which were 0.04 and 0.08, respectively,
although again the latter ratio was also based on a single sample.
Discussion
Plant species of Kawasi wetland
The number of species found in this study was lower than those
found in a similar study from Brazil (Carvalho et al. 2013). The
richness of species may be influenced by many factors. One of
them is the biogeographic distribution. Brazil, which is in the
Neotropical Biogeographic Region (Chambers et al. 2008)
had significantly higher species richness than the Australasian
Biogeographic Region in which Obi Island is located.
Besides the possibly of a naturally lower diversity in this
biogeographic region compared with that which encompasses
the Brazilian study, other explanations for the lower number of
species identified in this particular wetland include: (1) a limited
opportunity for other species to establish in this wetland because
seeds were not available as a consequence of the wetland being
surrounded by lowland fragmented secondary forests; (2) the fact
that the wetland experiences continuous inundation and anoxic
conditions in the saturated substrates would mean seeds of some
species, even if available, could not germinate well (Hammer
1997; Gibert et al. 2004); and (3) the current dominance of
existing macrophytes and their prolific regeneration from
rhizomes might limit the opportunities for other macrophyte
species to survive (Fig. 2).
Concentration of metals in plant tissue
It is inferred that the nickel and chromium in the water was derived
from the serpentine soil that is enriched in these elements, and
likely made more accessible as a result of the increased exposure
of these profiles due to mining activities seeking to extract nickel.
In this study, it was observed that the accumulation of chromium
and nickel in roots was higher than it was in shoots of all native
macrophytes. This was consistent with the findings of numerous
other studies examining metal accumulation in macrophytes,
wherein there are decreasing metal concentrations in the order
R. Amin et al.
of roots, stems, and leaves (Vymazal 2003; Vymazal and Krása
2003; Güleryüz et al. 2008; Gheju et al. 2009). However, we note
that the primary tissues of metal accumulation in macrophytes
do vary, both with species and type of contaminant (Baker 1981).
For example, Güleryüz et al. (2008) in their study investigating
heavy metals concentration in macrophytes showed that the
concentration of Zn in leaves was higher than in the stems
and roots of Xanthium strumarium, Urtica dioica, and Rumex
obtusifolius. Among these three species, R. obtusifolius
accumulated the highest concentrations of Zn, Mn, Ni and Cu
and the elemental concentrations themselves were higher in the
leaves than in the roots and stems.
Sheoran and Sheoran (2006) have suggested that there are
certain mechanisms in macrophytes that restrict the distribution of
elements such as chromium in plant tissues in order to protect the
plants from toxicity. As hexavalent chromium is toxic to plants, it
was suspected that for hexavalent chromium, the primary point of
separation and isolation of critical metabolic processes may be
through preferential accumulation in root tissue and minimal
transfer to the shoots. We also note that in four of the species
sampled, the shoot : root ratio of chromium was higher than that of
nickel, a result that may reflect a greater requirement by plants for
chromium as opposed to nickel since chromium, in its trivalent
form, is classified as a micronutrient (Blowes 2002; Tirez et al.
2007). This will be further studied in a later research project.
Acknowledgements
We thank Yopi Kalalo who accompanied and assisted the research activities,
the staff of the company’s Environmental Department, Trimegah Bangun
Persada, Gane Permai Sentosa, and especially Lusiana Kholifah and Hendrik
Supriyatna who made arrangements for travel, sample freight and secondary
data collection. We are very grateful to the management of the Harita Nickel
Group, especially Arif Perdanakusumah and Mufti Sodik, who allowed us to
conduct our study on Obi Island. Appreciation is also extended to Dr Antony
van der Ent and two anonymous reviewers for their comments on an earlier
version of this manuscript.
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