20 10 International Conference on Environmental Engineering and Applications (ICEEA 2010)

Heavy metals accumulation and tolerance in plants growing on ex-mining area, Bestari Jaya, Kuala Selangor, Peninsular Malaysia

Muhammad Ageel Ashraf; Mohd. Jamil Maah Department of Chemistry, Faculty of Science

University of Malaya 50603 Kuala Lumpur, Malaysia chemaqeel@gmail.com jamilmaah@gmail.com

Abstract-The degree of contamination by heavy metals in soil and transfer to plants has been studied. Specimens of plant species from five locations were sampled with their

corresponding soils. Thirty six plant species including two shallow water aquatic plants were identified. It was found that metal concentration in soil was highly variable while concentration of metals in plants directly depends on the

concentration of metals it was rooted. Bioconcentraion factor and translocation factor were calculated which show that Cyperus rotundus L. is a potential tin-hyperaccumulator plant, previously not reported in literature. Plant Species Imperata cylindrica, Lycopodium cernuum, Melastoma malabathricum,

Mimosa pudica Linn, Nelumbo nucifera, Phragmites australis

L., Pteris vittata L. and Salvinia molesta, were metal accumulator while Acacia podalyriaefoJia G. Don, Bulb

Vanisium, Dillenia reticulate King, Eugenia reinwardtiana, Evodia roxburghiania Hk. f. Clarke, Gleichenia Iinearis, Grewia erythrocarpa Ridl., Manihot esculenta Crantz,

Paspalum conjugatum Berguis, Passiflora suberosa, Saccharum officinarum, Stenochlaena palustris(Burm.) Bedd. and Vitis trifolia Linn. were tolerated plant species. All other studied plants were excluders. Identified plant species could be useful for revegetation and erosion control in metals­

contaminated ex-mining sites.

Keywords-Soil; roots; leaves; shoots; Cyperus rotundus L.; hyperaccumulator; hyper tolerant; remediation.

I. INTRODUCTION

Metals are non-biodegradable and therefore persist for long periods in aquatic as well as terrestrial environments. These metals may be transported through soils to reach groundwater or may be taken up by plants, including agricultural crops. Mine tailings are a major source of contaminants, mainly of heavy metals in water [1], soil [2] and biota [3] which has been extensively studied [4). The availability of metals in soil is a natural selection factor for plants that are capable of surviving or reproducing under high metal concentrations. Some plant species can grow in these severe conditions. Heavy metals cause oxidative stress in plants [5). Metal stress in plants affects photosynthesis, chlorophyll florescence and stomatal resistance. Copper inhibits photosynthesis and reproductive processes, lead reduces chlorophyll production, arsenic interferes with metabolic processes while zinc and tin stimulate the growth

978-1-4244-8621-2110/$26.00 © 2010 IEEE 267

Ismail Yusoff; Mohd. Mohamadreza Gharibreza Department of Geology, Faculty of Science

University of Malaya 50603 Kuala Lumpur, Malaysia ismaily70@um.edu.my

gharibreza4@yahoo.com

of leaves and shoots. Finally plant growth becomes limited or impossible. Plants can tolerate high heavy metals concentration from soil by two basic strategies [6]. The first strategy is called accumulation strategy where heavy metals are accumulated in plants at both high and low concentration in soil [7]. The second strategy is called exclusion strategy where transport of heavy metals in shoots and leaves are limited over a wide range of heavy metals concentration present in the soil. Based on these strategies some of the plants are called hyperaccumulator plants, as they can accumulate very high concentrations of metallic or metalloid elements in their aerial tissues, in excess of normal physiological requirements and of the levels found in most plant species [8). There are also some plant species called excluders that can restrict uptake and transport of elements between roots and shoots, maintaining low metal levels inside plant body over a wide range of external concentrations. More than 400 hypertolerant species have been identified [9], most of them associated with metal rich soils.

This research aimed to investigate polluted soils surrounding mining slag piles to identifY endemic excluder, accumulator and hyperaccumulator plant species or ecotypes, and in the medium term to evaluate their capability to remove heavy metals from the soil. This information could be useful to establish guidelines for the selection of adapted plant species that could be used for phytoremediation of these mining sites with low pH, high salinity and heavy metal concentration tolerances.

II. STUDY AREA

The present study was carried out in an old tin mininff area Bestari Jaya (Batang Berjuntai old name), located at 3 , 24' 40.41" N and 101024' 56.23" E, and is a part of district Kuala Selangor in Selangor state (Fig 1). Bestari Jaya was one of the most important tin mining sites in Selangor state and is now sand mining site. Bestari Jaya has a tropical, humid climate, with very little variations in temperature throughout the year. The average temperature of the area is 32 DC during day and 23 DC at night. An annual average rainfall of 2000 mm and 3000 mm with potential evaporation of 1600 mm per year [10).

2010 International Conference on Environmental Engineering and Applications (ICEEA 2010)

_,.O .... T C •• ","'N'HO ..... NO A .. IOI,CUI.TUIltAL LANa ...... N _AT ...• w ...... � ........ T

Figure I. District Kuala Selangor showing study area

Ill. MATERIAL AND METHODS

A. Sampling

Seven sampling sites were established (S I-S7) as shown in Fig 2. S 1 and S2 were located at downstream of catchment along sides of river Ayer Hitam, S3 in the reclaimed ex­mining area, S4, SS, S6 on mine dumps and S7 at a mineral processing tailing pond. These areas were selected based on topography, vegetation, and slope. Total seven 10 x 10 m plots were constructed. Plant samples (leaves, shoots, roots and flowers) were collected from every plant rooted in the sampling location. Each plant species sampled was divided into two groups. One group was preserved in S-S-S FAA solution (formic acid S%, acetic acid S%, methyl alcohol S% and distilled water 8S% by volume) for nomenclature. The nomenclature of the plant species is according to International code of botanical nomenclature [II). The second group of plant specie were each labeled with a numbered tag and were then pressed between sheets of paper in order to preserve them for laboratory analysis.

Figure 2. Sampling locations in the study area (Schematic diagram)

Soil samples (0--40 cm) were also collected at the same time and same place where the plants were sampled. Sampling location S7 includes a shallow pond area thus sediment core samples were collected along with corresponding plant species.

B. Soil Analysis

Analytical grade reagents were used throughout the experiment. The composite samples of soil and sediment were air-dried and milled so as to pass through a 2mm sieve,

268

homogenized prior to analysis. The pH was measured in a 1 :2.S soil/H20 suspension [12] using a waterproof pHiORP meter. Cation Exchange Capacity (CEC) was measured following the standard procedure [13] and the texture was analyzed by the hydrometer method [14]. Organic matter was determined by the Walkley and Black procedure [IS). For the analysis of metals homogenized soil samples were ashed in a muffle furnace at 400°C for I hour and were digested by microwave assisted acid digestion [16]. Solutions from digested soil samples were stored in 100mL high density polyethylene samples bottles at 4°C until analysis. Total metals in soil were measured in a sub-sample of O.S g soil inductively coupled plasma optical emission spectrometry ICP-OES (Varian) (Perkin Elmer AA Analyst). The methodology for total metal concentration in soil was referenced using the CRM027-0S0 Certified Material (Resource Technology Corporation, USA) and was analyzed concurrently with the soil samples.

IV. PLANT ANALYSIS

Plants samples were rinsed once with tap water, once with distilled water and finally twice with deionized water to remove adhering materials. Samples were digested by microwave assisted wet chemical digestion method [17], and analyzed by using inductively coupled plasma optical emission spectrometry ICP-OES (Varian) (Perkin Elmer AA Analyst). The methodology for total metal concentration in plants were referenced using CRM 281 [18] and was analyzed concurrently with the plant samples. Recoveries of metals by dry ash chemical destruction method of analysis were 99% for tin, 91% for arsenic, 9S% for copper, 90% for zinc and 93% for lead and the coefficient of variation was between 8% and 14% when analyzed in triplicate.

A. Determination of Translocation Factor (TF) and Bioconcentration Factor (BCF)

To analyze the total metal concentration in dry weight (DW) taken by the upper parts of the plants from ground level, a term was used called TF which is defined as [19]:

TF = Metal in above-ground DWI Metal in root DW Bioconcentration factor BCF, represents the ratio of

metal concentration DW in the plant to the metal concentration DW in the soil. BCF is expressed as [20]:

BCF= Metal in Whole plant DW IMetal in soil DW

V. RESULTS AND DISCUSSION

A. Characterization of Soil

Soil pH values ranged from acidic to neutral (4.8-7.2). The pH was acidic in the location (S4--S6), while it was slightly acidic at (SI-S3) and was neutral at (S7).The carbonate percentages were in a broad range and the organic matter values were less than 10%. The CEC represents the ability of the soils to absorb or release cations, and consequently is an important parameter in sites contaminated by heavy metals. Organic matter and clay minerals are responsible for the CEC. CEC ranged from low 17.81 to high 26.98 cmollkg. pH and ECs are the most important factors [21] because under acidic conditions the tailings matrix will

2010 International Conference on Environmental Engineering and Applications (ICEEA 2010)

dissolve more salts [22). At all sampling locations, the soil showed a sandy texture. Sandy substrates generally present oxidizing conditions however, in this case, the water­saturation state of soils and the flooding of sediments explained the reducing environment. In the study area analyzed metal contents in soils were highly variable and also found to be high depending on the mine and type of ore [58]. At sampling location (SI-S3) in the reclaimed and river side, metals concentration were averaged from 13 to 89, 11 to 47, 18 to 71, 8 to 91, 123 to 271 mg/kg DW for lead, copper, zinc, arsenic and tin respectively while sampling location (S4-S7) in the mine dumps and the tailing ponds showed mean values of 541 to 3589, 761 to 2781, 638 to 3698, 239 to 2956, 1896 to 6453 mg/kg DW for lead, copper, zinc, arsenic and tin respectively.

B. Identification and Classification of Plants

Thirty six plant species were identified in the study area (Table 2) that belong to different plant families such as Blechnaceae (I), Casuarinaceae (1), Compositae (1), Cucurbitaceae (1), Cyperaceae (1), Dilleniaceae (2), Euphorbiaceae (4), Fabaceae (3), Gleicheniaceae (1), Gramineae (1), Lycopodiaceae (1), Malvaceae (1), Melastomataceae (1), Mimosaceae (I), Myrtaceae (1), Nelumbonaceae (1), Nepenthaceae (1), Orchidaceae (1), Passifloraceae (I), Poaceae (3), Pteridaceae (1), Rutaceae (I), Salviniaceae (I), Schizaeaceae (1), Tiliaceae (1), Ulmaceae (I), Verbenaceae (I) and Vitaceae (1) largely as a result of the sampling criteria. Some of the identified species, such as Acacia podalyriaefolia G. Don, Antidesma ghaesembilla Gaertn. , Evodia roxburghiana Hk. j, Imperata cylinderica, Lygodium mycrophyllum (Cav. ) R. Br. , Melastoma malabathricum L. , Phragmites australis L. , Stenochlaena palustris(Burm. ) Bedd., were sampled from three or more locations in the study area (Table I).

Plant species were categorized according to their TF values into four groups (Table I). First group contain hyperaccumulator plants with TF values above ten (Table 1, 1). Only Cyperus rotundus L. follow the criteria for hyperaccumulator. Total 1990.44 mg/kg of metal concentration was found in roots, shoots, leaves and flowers which show that this plant has a potential to hyperaccumulate tin metal, previously not reported in literature. Second group contain hpertolerant plants with TF values above one but below ten (Table I, 2). Results in (Table I, 2) indicate that these plants adopted an accumulation strategy with regard to different studied metals. Eight plant species were identified as dominant hypertolerant species that includes Imperata cylindrica, Lycopodium cernuum, Melastoma malabathricum, Mimosa pudica Linn, Nelumbo nucifera, Phragmites australis L. , Pteris vittata L. and Salvinia molesta (Table 1). Third group contains tolerant plants with TF values less then one. These include Acacia podalyriaefolia G. Don, Bulb Vanisium, Dillenia reticulate King, Eugenia reinwardtiana, Evodia roxburghiania Hk. j Clarke, Gleichenia linearis, Grewia erythrocarpa Ridl. , Manihot esculenta Crantz, Paspalum conjugatum Berguis, Passiflora suberosa, Saccharum officinarum, Stenochlaena palustris(Burm. ) Bedd. and Vitis trifolia Linn. (Table I).

269

These results supported the idea that plants have adopted an exclusion strategy. The fourth group, categorized as excluders, as these plants can grow in heavy-metal polluted soils without accumulating significant quantities [6]. These include Casuarina equisetifolia Forest, Eupatorium odoratum L, Momordica balsamina L. , Tetracera indica Merr. , Antidesma ghaesembilla Gaertn. , Bridelia monoica (Lour. ) Merr. , Mallotus paniculatus (lam. ) Mull. Arg ,Crotalaria retusa Linn, Desmodium gangeticum L. , Hibiscus tiliaceus L. , Nepenthes gracillis, Lygodium mycrophyllum (Cav. ) R. Br. , Trema orientalis L. and Stachytarpheta indica Vahl. These plant species have TF values < 0.1. Possibly these plants use mechanism that avoids excessive uptake and metal is absorbed and translocated only in required or non-toxic quantities. These plants are not useful for phytoextraction treatments.

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20 I 0 International Conference on Environmental Engineering and Applications (ICEEA 20 I 0)

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Figure 3. Dominant plant species with heavy metals concentration

Lead occurred at the highest concentrations (Table 2; Fig. 3a) in five species exactly in Cyperus rotundus L, Imperata cylinderica, Lycopodium cernuum, Phragmites australis L. , and Salvinia molesta where we obtained values around 897.08 mg/kg lead in Imperata cylinderica. The lowest values were recorded in Lycopodium cernuum with around 94.11 mglkg lead. Regarding the plant accumulation of Copper (Table 2; Fig. 3b), three species had significantly higher concentrations, namely Imperata cylinderica, Phragmites australis L and Nelumbo Nucifera, with 684.97 mg/kg, 717.49 mg/kg and 1125.08 mglkg, respectively. The lowest values of copper were obtained for Cyperus rotundus L, Lycopodium cernuum, Melastoma malabathricum L. , Mimosa pudica Linn. , Pteris vittata L. and Salvinia molesta with less than 64.11 mg/kg copper in Lycopodium cernuum. Highest concentration of zinc 731.92 mglkg was found in Imperata cylinderica (Table 2; Fig. 3c) whereas lowest concentration 133.45 mglkg was found in Pteris vittata L. Arsenic occurred high in Pteris vittata L. and Nelumbo Nucifera where it was 929.53 mg/kg in Nelumbo Nucifera (Table 2; Fig. 3d) while lowest 79.02 mg/kg was in Melastoma malabathricum L. Tin concentration was found significantly higher in four plant species namely Cyperus rotundus L, Imperata cylinderica, Melastoma malabathricum L.and Pteris vittata L. The highest concentration of tin 1990.44 mg/kg was found in Cyperus rotundus L with a potential of being hyperaccumulator plant (Table 2; Fig. 3e) while the lowest 310.00 mglkg was found in Phragmites australis L. Bioconcentration factor BF values [60] for the group first and second plants were from 0.2181 to 0.7638 for lead, 0.2670 to 1.5705 for copper, 0.1327 to 0.7258 for

270

zinc, 0.3014 to 0.5179 for arsenic and 0.1715 to 0.4022 for tin (Table 3). These high values indicate that plant species are suitable for phytoextraction and follow the criteria of hypertolerants while fourth group plants have values in range (0.0091-0.067) which show that these plants can be suited to soil stabilization because there is less risk of metals entering the food chain if the plants are consumed. The results obtained thus confirmed Cyperus rotundus L. as hyperaccumulator plant previo�;,��,.not reported in literature.

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TABLE II HEAVY METALS CONCENTRATION IN DOMINANT PLANTS

VI. CONCLUSION

Metals transport in surface waters and leaching through the soil profile may result in the contamination of consumable water through mining catchment as it flows into the native river. Meanwhile the growth of identified plant species can help to reduce the metal contaminants upto large extent. There is about 113,700 hectors of tin tailings throughout the Peninsular Malaysia. Identification of new plant specie as a potential tin hyperaccumulator is a major break through but still needs more exploration to make the specie feasible for growth in the tin tailings areas.

Figure 4. Cyperus rotundus L. as a tin hyper accumulator plant

20 10 International Conference on Environmental Engineering and Applications (ICEEA 2010)

ACKNOWLEDGMENT

The work reported in this paper was carried out in Analytical Laboratory, Department of Chemistry, University of Malaya, Kuala Lumpur, Malaysia through UM Research Grant vide no. PS35512009C. Thanks also to the Ministry of Higher Education Malaysia (MOHE) for their financial support.

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