Environ Monit Assess (2012) 184:51395150 DOI 10.

1007/s10661-011-2328-8

A study on the waste metal remediation using floriculture at East Calcutta Wetlands, a Ramsar site in India
Soumya Chatterjee & Lokendra Singh & Buddhadeb Chattopadhyay & Siddhartha Datta & S. K. Mukhopadhyay

Received: 12 December 2010 / Accepted: 29 August 2011 / Published online: 30 September 2011 # Springer Science+Business Media B.V. 2011

Abstract Use of specific plant species in remediation of heavy metal-contaminated soil and water was a promising eco-friendly technology. The present study indicated the possibilities of phytoremediation of metal-contaminated (namely Ca, Cr, Mn, Fe, Cu, Zn, and Pb) soil by using plant species important for floriculture of East Calcutta Wetlands, a Ramsar site at the eastern fringe of Calcutta city. Plant species like sunflower (Helianthus annuus), marigold (Tagetes patula), and cocks comb (Celocia cristata) grew on soil contaminated by industrial sludge and irrigated regularly with wastewater accumulated different metals in different plant parts in varied concentrations. Pot culture study in the laboratory setup was also done to
S. Chatterjee (*) : L. Singh Defence Research Laboratory, Post Bag No. 02, Tezpur 784001, Assam, India e-mail: drlsoumya@gmail.com B. Chattopadhyay Government College of Engineering and Leather Technology, LB, III, Salt Lake, Kolkata 700 098, India S. Datta Department of Chemical Engineering, Jadavpur University, Kolkata 700 032, India S. K. Mukhopadhyay Hooghly Mohsin College, Chinsurah 712 101, West Bengal, India

ascertain the efficiency of these plants for ameliorating contaminated soil. It was found that general accumulation patterns of metals concerned in different plant parts were root > leaf > stem > flower. This work indicated the importance of cultivation of economically important, non-edible, ornamental plant species as an alternative cost-effective practice to remediate heavily contaminated farmlands of East Calcutta Wetlands. Keywords Phytoremediation . Floriculture . Heavy metals . East Calcutta Wetlands . Ramsar site . Agro-soil

Introduction Environmental pollution with waste metals continues to be an important concern worldwide. Shipping or dumping of untreated industrial wastes are considered as serious factors for disturbing natural ecosystems, as they carry elevated concentrations of different elements into the area contaminating sediments, soils, and water. Further, the pollution constitutes a potential cause of health risks for plants, humans, and other organisms in the food web (Merdy et al. 2006). Though elements like Ca, Cu, Zn, Mn, and Fe are essential to biological systems at optimal levels, they are toxic when present in excess. For the past few years, there is a growing concern over eco-friendly, cost-effective technologies for remediation of contaminated areas (McIntyre 2003). Some plants can persist in heavy metalcontaminated areas and had the ability to accumulate

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different metals within their bodies. One of the most appealing attributes of phytoremediation is that when the appropriate plants are used, cleanup can be ensured in sites that are contaminated with heavy metals contaminants. To date, several plant species have been identified that can be used for removing Cu, Cd, Co, Mn, Ni, Zn, etc. from soils containing contaminant levels of those elements. Notable published reports on this issue include Prasad (1995), Das et al. (1998), Lombi et al. (2001), Garbisu et al. (2002), Lasat (2002), McIntyre (2003), Singh et al. (2003), Lu et al. (2004), and Mendez and Maier (2008). However, no phytoremedial measures by the use of ornamental plants which are economically important have been attempted in the East Calcutta Wetlands, a Ramsar site (the wetlands of international importance, known as Ramsar Convention, is an international treaty for the conservation and wise use of wetlands and their resources) on the eastern fringe of Kolkata city, India. This unique and biggest urban ecosystem, which is the part of mature delta of river Ganges, is being vitiated by composite tannery wastewater from at least 538 tanneries mixed with the effluents of around 5,500 other industries and wastewater of Kolkata city. The composite effluent (around 50,000 m3 day1) is discharged into the wetland ecosystem without any pretreatment. The wastewater flows down through a web of canals traversing the ecosystem and is finally discharged in Kultigong, nearly 40 km away from the city. On this journey, the wastewater is productively utilized to irrigate agricultural farms and in pisciculture tanks (Chatterjee et al. 2006, 2009). Such uses over the years have loaded the soil, sediments, and water with high levels of various elements. In the present study, phytoremediation of the waste elements in the soils of agricultural land with the help of certain plant species especially used for floriculture, namely sunflower (Helianthus annuus), marigold (Tagetes patula), and cocks comb (Celocia cristata) have been investigated as they would possibly show us a way to clean up operation which is economical, non-intrusive, esthetically pleasing, and involves low-cost technology.

Materials and methods The samples were collected from different fields of East Calcutta Wetlands area (latitude 22332240 N, longitude 88258835 E) where the composite

tannery effluent was used to irrigate the fields regularly. Three common plant species important from floricultural standpoint, namely sunflower (H. annuus), marigold (T. patula), and cocks comb (C. cristata), were collected from ten randomly selected sites. Ten plants of each species from chosen fields were collected with wet weight ranged from 800 to 900 g for sunflower, 200300 g for marigold, and 900 1,050 g for cocks comb. Soil samples were collected (using a plastic shovel and kept in uncontaminated wide-mouthed glass vials of 250 ml capacity with caps) from the sites before the cultivation and from the root zones of the collected plants for examination of elemental concentrations. Laboratory-based pot culture experimentation had also been carried out for estimation the efficiency of selected plants (sunflower, marigold, and cocks comb) in bioremediation of contaminated soil. Fifteen pots (each with an effective volume of 9,82020 cm3), five replicates for each, were taken and filled with the same contaminated soil collected from our study site, the East Calcutta Wetland area. The metal concentration of the soil sample, used to fill up the pots, was also estimated as described below. Saplings of sunflower, marigold, and cocks comb were planted after weighing (sapling wet weight ranged from 1.60.5 g for sunflower, 1.10.6 g for marigold, and 2.40.8 g for cocks comb) and allowed to grow until the blooming period was over, the usual period of exposure of the selected plants in the fields. No fertilizer or pesticide was used. Only distilled water was used for watering purposes. Soil samples were also analyzed for the elements before planting and after uprooting to study efficacy of the plants in metal uptake from the soil. Biomass increment of the plants was also recorded (wet weight ranged from 70456.2, 19222.1, and 81139.2 g for sunflower, marigold, and cocks comb, respectively) over a period of 90 150 days, depending on the period of maturity and flowering of the concerned plant . Deracinated plant samples were thoroughly cleaned with Millipore water (18 M water from a Millipore water purification unit) and separated into root, stem, leaf, and flower. Root parts were cleaned very carefully by repeated washing using Millipore water to avoid erroneous result. These were then cut into small pieces and dried in an air oven slowly at a 90C. Dried grounded plant parts were digested following standard methods (Eaton et al. 1995). Briefly, 1.00.05-g dried grounded plant parts were

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taken inside a 100-ml Borosil beaker. Ten milliliters of concentrated HNO3 (E. Merck) was added, swirled, and the mixture was kept for 1012 h with occasional stirring. Few porous beads were added and the mixture was heated (with caution) over a hot plate gently until the escape of NO2 fume ceased. The beaker was cooled to room temperature. A mixture of 10 ml each of HNO3 (conc.) and HClO4 (70% pure) was added, and the mixture was boiled on a hot plate (with caution) until the volume of the suspension reduced approximately within 35 ml. This mixture was then cooled to room temperature. The solution was then transferred into a 50-ml volumetric flask, and then Millipore water was added up to the mark. The extracts thus prepared were stored inside a refrigerator for determination of Ca, Cr, Mn, Fe, Cu, Zn, and Pb by AAS following standard methods (Eaton et al. 1995). Soil samples were dried, ground to powder, and sieved for extraction of total Ca, Cr, Mn, Fe, Zn, Cu, and Pb from dry ashes following Isaac and Kerber (1971) and Welz and Sperling (1999). About 1.0 0.05-g dried and grounded soil sample was placed inside a silicone crucible and ignited in a Muffle furnace at 500C for 3 h. The ignited mass was cooled inside a desiccator and transferred into a 100-ml Borosil beaker. Inside the beaker were added 10 ml 1:1 HCl and the suspension swirled. The suspension was kept inside a thermostat-controlled water bath kept within the range 70 80C for 1 h. The supernatant was decanted and kept inside a 250-ml volumetric flask with stopper. This contains mostly alkaline earth metals. To the residue in the beaker, 10 ml each of HCl (conc.) and HClO4 (conc., 70% pure, E. Merck India) and few porous beads were added and evaporated to complete dryness over a hot plate. This process was repeated as such once more and further with half the amount of acid mixture once. The dried residue was dissolved completely by using minimum amount of conc. HCl. This solution was then transferred to the same volumetric flask where previous extract containing alkaline earth metal extracts was stored. The flask were then made up to the mark by Millipore water and stored inside a refrigerator. This extracts were analyzed for metals. Metals, viz. Ca, Cr, Mn, Fe, Cu, Zn, and Pb were detected by atomic absorption spectrophotometer (Perkin-Elmer AAnalyst 100) using specific hollow cathode lamp in default condition by flame absorption

mode. Each time the concentration was determined using non-linear calibration with three replicates each and 3.0 s integration times. The mean concentration, standard deviation, and relative standard deviation were determined. The detection limit of the instrument for the element mentioned here was 0.01. Standards recommended by Perkin-Elmer were used for both checking the sensitivity of the instrument and calibration. Reference materials (water: SRM 1643d, estuarine sediments: 1646a, apple leaves: SRM 1515) procured from National Institute of Standards and Technology were also prepared along with the sample in the same way mentioned above. All gravimetric works, reagent, and standard preparations were made using Mettler AE 240 monopan electronic balance. pH and the reagent factors were determined using Mettler Autotitrator DL 25. Analytical results were evaluated using SPSS (version 14.0). The standard deviation values were calculated, and the representative graphs were made using Origin (version 6.1). ANOVA (F test) was performed to compare the different study sites. Other statistical analyses like PCA, determination of correlation matrix, and clusters were performed using SPSS (version 14.0) to evaluate further the different components and parameters of the dataset.

Results Elemental uptake by plants in the field Elements in sunflower, marigold, and cocks comb of contaminated areas were studied for different metal accumulations (Fig. 1ad). It was found that general accumulation pattern of metals concerned in different plant parts were root > leaf > stem > flower. Wide variation of the concentration level in different parts of the plants had been recorded. Ca concentrations of roots of all three plants were high than that of other aerial parts. The highest concentration of Ca was found in the root of cock s comb (11,024 181.7 mg1 kg DW) followed by marigold (10,234 195.6 mg 1 kg DW) and sunflower (8,562 159.6 mg1 kg DW). Leaf also accumulated considerable amount of Ca with the highest accumulation observed in marigold (2,69856.1 mg1 kg DW). The Cr concentration in the root of cocks comb (84 5.6 mg1 kg DW) was recorded the highest followed

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a
12000 11000 10000 9000 8000 7000 6000 5000 4000 450 400 350 300 250 200 150 100 50 0

b
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sunflower marigold cock's comb

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mg/kg DW
Ca Cr Mn Fe Cu Root (field) Zn Pb

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sunflower marigold cock's comb

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Ca

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Fe Leaf (field)

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Ca

Cr

Mn

Fe

Cu

Zn

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Flower (field)

Fig. 1 ad Waste metals in different plant parts (root, stem, leaf, and flower) of sunflower, marigold, and cocks comb collected from fields. a Metals in roots, b metals in stem, c metals in leaf, d metals in flower

by root of sunflower (5612.8 mg1 kg DW). Among aerial parts, maximum accumulation of Cr was in leaf with the highest in cocks comb (39 3.2 mg kg1 DW) and the lowest in flower of sunflower (62.1 mg kg1 DW). The concentrations of Mn in root (7119.5 mg kg1 DW) and leaf (54 13.1 mg kg1 DW) were recoded the highest in sunflower among others of the respective parts. Similarly, cocks comb was recorded as major accumulator of Fe with the highest concentrations in all parts among other plants. The pattern of concentration of Fe in cocks comb was as follows: root (4,732467.2 mg kg1 DW) > leaf (1,718 225.1 mg kg1 DW) > stem (88926.3 mg kg1 DW) >

flower (284.5 mg kg1 DW). Average Cu accumulation in sunflower was the highest (32 mg kg1 DW) followed by marigold (22 mg kg1 DW) and cocks comb (16 mg kg1 DW). Interestingly, the highest concentration of Cu was found in the root of sunflower (489.1 mg kg1 DW), followed by the flower (29 4.2 mg kg1 DW). Zn accumulation in the root of sunflower was the highest (46924.4 mg kg1 DW), followed by cocks comb (43748.2 mg kg1 DW). Significant amount of accumulation of Zn in leaf of the plants was also found with the highest concentration in cocks comb (36538.9 mg kg1 DW), followed by sunflower (35545.3 mg kg1 DW). Pb content in the roots of sunflower, marigold, and cocks comb were

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47.910.25, 434.2, and 7212.8 mg kg1 DW, respectively. The concentration of Pb in the leaf of sunflower (447.7 mg kg1 DW) and cocks comb (425.8 mg kg1 DW) was almost similar. Interestingly, the highest concentration of Pb was found in >flower of marigold (255.7 mg kg1 DW) and the lowest in flower of cocks comb (101.2 mg kg1 DW). Changes in metal concentrations in field soil It had been found from our study that, on the average, the root level soils of three selected plants showed significant decrease in elemental concentration levels when compared to the concentrations before the cultivation was started (Fig. 2). Cr, a known contaminant of tannery wastes, was present in the field soil in a concentration level of 64629.3 mg kg1 DW, compared to that of cultured soil 52317.8 mg kg1 DW. In contaminated soil, the concentrations of Ca (25,419 566.1 mg kg1 DW) and Fe (2,116 372.7 mg kg1 DW) were very high; however, after cultivation, the field soils of root zones were estimated as 22,394310.2 mg kg1 DW for Ca and 17,870209.3 mg kg1 DW for Fe. The non-essential toxic element Pb was also reduced in the soil after cultivation of the plants. The initial concentration of the field soil was 985.0 mg kg1 DW for Pb, while that of after cultivation was 615.8 mg kg1 DW. Overall, the reductions in the concentrations of
Field soil before cultivation Root soil after cultivation of flowering plants
28000.0 26000.0 24000.0 22000.0 20000.0

different metals in the root zone soils after the floriculture season were 11.9% for Ca, 19% for Cr, 13.8% for Mn, 15.4% for Fe, 22.2% for Cu, 25.2% for Zn, and 27.9% for Pb in respect to that of uncultivated contaminated soils. Pot culture study on metal uptake by plants Pot culture experimentation in the laboratory of the same three species, sunflower, marigold, and cocks comb showed almost similar pattern of accumulation of different heavy metals in their body parts with the highest concentrations in roots (Fig. 3). Average Ca accumulation was recorded the highest in cocks comb (2,154 mg kg1 DW) followed by sunflower (1,773 mg kg1 DW) and marigold (1,729 mg kg1 DW) with the highest accumulation in root of cocks comb (5,715123.8 mg kg1 DW) and the lowest in marigold flower (35927.9 mg kg1 DW). Similarly, average Cr accumulation was recorded the highest in cocks comb (28.9 mg kg 1 DW) followed by sunflower (20.7 mg kg1 DW) and marigold (15.8 mg kg1 DW) with the highest accumulation in root of cocks comb (595.4 mg kg1 DW) and the lowest in marigold flower (50.2 mg kg1 DW). However, average Mn concentration was recorded the highest in sunflower (39.7 mg kg1 DW) with considerable amount in root (65 3.5 mg kg1 DW) followed by leaf (59 7.1 mg kg1 DW), flower (242.7 mg kg1 DW), and stem (104.2 mg kg1 DW). Concentration levels of Fe in all parts of the cocks comb were documented higher than other two plants. The average concentration levels
2400 2200 2000 1800 1600 1400 1200 1000 800 600 400 200 45 40 35 30 25 20 15 10 5 0 Ca Cr Mn Fe Cu Zn Pb

Sunflower Marigold Cock's comb

Conc (ppm) dw

18000.0

600.0 500.0 400.0 300.0 200.0 100.0 Ca Cr Mn Fe Cu Zn Pb

Metals in field soil

mg/Kg DW

16000.0

Metals in Pot Plants

Fig. 2 Metal profile in the uncultivated field soil (n =10) and the root soil (n =10) collected from different fields, grown-up with marigold, sunflower, and cocks comb for cultivation (concentration in milligrams per kilogram dry weight)

Fig. 3 Metal accumulation profile in sunflower, marigold, and cocks comb cultivated in pots (n =5 for each plant, concentration in milligrams per kilogram dry weight)

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of Fe in plants were cocks comb (1,752 mg kg1 DW) > sunflower (1,421 mg kg 1 DW) > marigold (1,389 mg kg1 DW). Cu concentration was recorded the highest in sunflower (24.7 mg kg1 DW) with an interesting accumulation pattern within their body. The maximum accumulation was found in root (35 5.4 mg kg 1 DW) followed by flower (23 1.6 mg kg1 DW), stem (222.9 mg kg1 DW), and leaf (184.1 mg kg1 DW). Average concentration of Zn was recorded the highest in cocks comb (204 mg kg1 DW) followed by sunflower (199 mg kg1 DW) and marigold (163.0 mg kg1 DW). Average mean Pb concentration in sunflower and cocks comb was almost similar (24.4 and 24.1 mg kg1 DW, respectively) with slight higher concentration in marigold (28.9 mg kg1 DW). Interestingly, the concentration pattern of Pb in leaf part of all three species was recorded almost similar with 27 6.2 mg kg1 DW for sunflower, 26 5.3 mg kg1 DW for marigold, and 261.8 mg kg1 DW for cocks comb. Changes in metal concentrations in pot soil Pot soils were analyzed before plantation and after uprooting of the respective plants. The mean concentration levels of different metals in the pot soil before plantation were 19,561 mg kg 1 DW for Ca, 235 mg kg1 DW for Cr, 474 mg kg1 DW for Mn, 15,214 mg kg1 DW for Fe, 75 mg kg1 DW for Cu, 552 mg kg1 DW for Zn, and 60 mg kg1 DW for Pb. Figure 4 depicted the change in the concentration level in the pot soil after the planting of three selected plant species, sunflower, marigold, and cocks comb. Variation in the waste metal remediation was recorded. After cultivation, the Ca concentrations in the soils of sunflower, marigold, and cocks comb were 13,424251.7, 13,958521.9, and 13,129 897.6 mg kg1 DW. Similarly, following the cultivation, Cr concentrations in the soils of sunflower, marigold, and cocks comb were 15813.9, 186 19.6, and 152.519.17 mg kg1 DW, respectively. Percentage reduction in pot soil for Cr was cocks comb (35.1%) > sunflower (32.5%) > marigold (20.9%). It had been found that percent decline of Fe in all three plants was almost similar (around 17%) with final concentration in pot soils of sunflower, marigold, and cocks comb of 12,618529.2, 12,490243.6, and 12,536244.2 mg kg1 DW, respectively. Cu concentration in pot soil after cultivation also showed

significant reduction in the concentration level. Sunflower and cocks comb soil showed almost similar decrease (31.1% and 31.3%, respectively) and marigold 34.7% in Cu concentration level. The final concentrations of Zn in the pot soils for sunflower (417.5 35.38 mg kg1 DW), marigold (455.5 29.68 mg kg1 DW), and cocks comb (424.7 22.9 mg kg1 DW) showed an effective percentage reduction of 24.4%, 17.5%, and 23.1% respectively. Interestingly, it may be indicated from our data that the pot soils of sunflower and cocks comb showed almost similar reduction in the concentration levels of the particular metals, viz. Zn, Pb, and Cu. Among all other metals, Pb was found to be the highest removed from the soil with an effective percentage reduction of 71% in sunflower, 49% in marigold, and 72% in cocks comb. Least effective reduction in the concentration level in pot soil was found in case of Mn with 5.7%, 7.8%, and 11% in sunflower, marigold, and cocks comb, respectively. Statistical analysis The factor analyses using principal component analysis (Varimax with Kaiser normalization) were performed (Table 1), and the range of factor loading (FL) between 0.25 and 1.0 for each component was emphasized. Factor analysis reveals the importance of heavy metals in field soil, and pot soil with the first component as per highest factor loading follows the sequence for field experiment: Ca (FL=0.857) > Fe (FL=0.838) > Pb (FL=0.685) > Cr (FL=0.619) > Zn (FL=0.479) and for pot culture experiment: Ca (FL=0.886) > Pb (FL= 0.806) > Cr (FL=0.805) > Fe (FL=0.768) > Zn (FL= 0.605). In the second component, Cu (FL=0.778) for field and Mn (FL=0.876) for pot culture while in the third component Cr (FL=0.715) for field and Cu (FL= 0.828) for pot culture were evident. Therefore, it is interesting to note that abundant bioactive metals, namely Ca and Fe, are strongly interacting and/or influencing the mobility and partitioning of the other unwanted metal ions, namely Pb and Cr, available in the ambient environment. Dendrogram constructed (Fig. 5) on the relation between the different plant parts that accumulated unwanted metals showed a distinct cluster distribution. Roots of all the three plant species formed a distinct cluster while the leaves were in a separate distinct cluster. Stems and flowers of the three plant species studied were illustrated a further

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Ca
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Pr. Pot soil SFSoil MGSoil

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500 400

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Pr. Pot soil SFSoil MGSoil CC Soil

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Pot soil

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Pr. Pot soil SFSoil MGSoil CC Soil

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Fig. 4 Concentration levels of waste metals in pot soils before cultivation (Pr Pot Soil) and after cultivation of sunflower (SF Soil), marigold (MG Soil), and cocks comb (CC Soil) (concentration in milligrams per kilogram dry weight)

distinct cluster. A characteristic pattern in the plantparts-wise preference in metal accumulation could be envisaged from the plot. Moreover, a very strong

positive correlation (r =0.991, at p <0.05) between the initial and the final metal concentrations in soils and plants indicated that plant uptake was much dependent

5146 Table 1 Component matrix for principal component analysis with mean data for metal accumulations in different parts of the plants collected from field trials and pot culture Metals in plant parts Component 1 Ca in plants from field samples Ca in plants from pot samples Cr in plants from field samples Cr in plants from pot samples Mn in plants from field samples Mn in plants from pot samples Fe in plants from field samples Fe in plants from pot samples Cu in plants from field samples Cu in plants from pot samples Zn in plants from field samples Zn in plants from pot samples Pb in plants from field samples Pb in plants from pot samples 0.857 0.886 0.619 0.805 0.296 0.270 0.838 0.768 0.377 0.113 0.479 0.605 0.695 0.806 2 0.438 0.358 0.091 0.133 0.660 0.876 0.417 0.394 0.778 0.234 0.630 0.573 0.306 0.421 3 0.229 0.246 0.715 0.544 0.585 0.236 0.298 0.480 0.100 0.828 0.571 0.476 0.553 0.038

Environ Monit Assess (2012) 184:51395150

Extraction method: principal component analysis with three components extracted. Rotation method: Varimax with Kaiser normalization

on the available concentrations in the ambient environment (Fig. 6).

Discussion The waste metal-contaminated soil and irrigating wastewater were the sources of accumulation of
Fig. 5 Relation between the plant parts that accumulated unwanted metals at the contaminated East Calcutta Wetlands. 1=sunflower root, 2=marigold root, 3= cocks comb root, 4=sunflower stem, 5=marigold stem, 6= cocks comb stem, 7=sunflower leaf, 8=marigold leaf, 9= cocks comb leaf, 10=sunflower flower, 11=marigold flower, 12= cocks comb flower

different elements in the plants grown in the concerned area. The uninhibited entry of elements throughout the year in soils through watering by contaminated water and the use of sewage sludge were alarming because metals once accumulated in the soil were usually hard to remove (Smith et al. 1996). Most of the elements under investigation, like Ca, Fe, Mn, Cu, and Zn, were known to be essential for higher plants. However, metals such as Cr and Pb did not play any significant role in the plant physiology (Marschner 1995). Plants growing in a polluted environment can accumulate the toxic metals at high concentration causing serious risk to the consumers of the ecosystem (Vousta et al. 1996; Singh et al. 2003; Bertrand and Poirier 2005; Chatterjee et al. 2007). In the present study, three non-edible plants important from floricultural standpoint, namely sunflower, marigold, and cocks comb, were found to be good accumulator of different elements. Concentration of different elements in different parts followed a trend: roots> leaves> stem> flower. Roots and leaves were the dominant sites for metal accumulation of plants grown in polluted environment as was also supported by earlier workers (Anikwe and Nwobodo 2002). The effects of any element on the plant depended on qualitative and quantitative properties and concentration of other elements and organic substances. However, metal-specific accumulation by three different plant species was evident from our study. Fe was observed to be present in higher concentration in the root possibly because younger tissues were dependent on a continuous Fe supply in

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Fig. 6 Regression plot between the relation of initial and final metal concentrations in field soil, pot soil, and different plants cultivated in fields and pots

the xylem (Mengel and Kirkby 1987). Zn associated with Cu acts as prosthetic groups in various enzymes. Gradual translocation of Zn took place from the root to the leaves for obvious metabolic purposes (Xian 1989; Okoronkwo et al. 2005). Thus, higher Zn and Cu accumulation without apparent toxic effect in all of these plants studied was noted in stem and leaf possibly because of the higher enzymatic activity in them (Wainwright and Woolhouse 1975). Metal metal interactions and competition and availability of organic substances at the rhizospheric zone of the plant played an important role in sequestration and bioaccumulation (Birge et al. 2000; Madejon et al. 2003). For example, metal toxicity of Cu to algae became more pronounced if the organisms were stressed by Mn limitation (Sunda and Huntsman 1983). Similarly, recently, Alam et al. (2006) reported the alleviation of Mn phytotoxicity with Ca. Other elements like Fe, Mn, Cu, and Zn were essential at low concentrations but were toxic at higher levels (Kitao et al. 1997; Doncheva et al. 2006; Paschke et al. 2006). Bioavailability of such nutrient elements was also affected by clay content, soil permeability, moisture

content, etc. (Hering and Kraemer 1998). However, organometallic Cu complexes played a key role in regulating Cu mobility than that of other micronutrient cations like Fe and Mn (Lidon and Henriques 1992). The higher concentration of Mn in leaves of all the plants collected from both the fields and pots might be due to bioactive role of the element in the particular organ. It was interesting to note from the present study that the uptake and translocation of Cr was found low in contrast to Pb though the concentration level of Cr in the soil was much higher than that of Pb. This was presumably because the soil Cr was largely unavailable to plants as it (Cr3+) bound tenaciously to negatively charged sites of clay and organic matters (Banks et al. 2006). Pb was toxic to plants because it mimics the metabolic behavior of Ca and inhibits many enzymatic reactions (Clarkson 1993). Organic lead was extremely mobile in soil and was taken up by plants much more readily than Pb2+ (Schwab et al. 2005). Plants distribute metals internally in many different ways. Accumulation of Ca in all taxonomic levels of photosynthetic plants was pronounced by the forma-

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tion of substantial amount of calcium oxalate crystals within their parts, which was helpful for high-capacity calcium (Ca) regulation in Ca-rich environment (Franceschi and Nakata 2005). A mechanism of tolerance and accumulation in some plants apparently involves binding potentially toxic metals at cell walls of roots and leaves, away from sensitive sites within cell or storing them in vacuolar compartments (Memon et al. 1979, 2001). Adaptation of plants to accumulate metals in high concentrations without apparent physiological dysfunction involved methods including chelation and/or methylation of the metals by producing phytochelatins (Cobbett 2000) and metallothionein (Rauser and Curvetto 1980; Robinson et al. 1993, 1997), major metalloproteins in the body that reduces its toxicity. In the present study, the chosen plants important from floricultural standpoint were characteristically tolerant and showed ability to produce huge biomass in metal-rich soils, absorbing heavy metals into the roots, and transferred metals to the aboveground parts exhibiting all enviable features for effective phytoremediation.

market in the areas. Present study indicated that sunflower, marigold, and cocks comb, usual cultivable species of the areas, were able to take up metals from the contaminated sites. It could, therefore, be suggested to culture these common economically important flowering plants for a season or two, making use of the heavily contaminated agro-soil, to ameliorate the hazardous metal load of the sites. Additionally, raw wastewater of the canal should not be used for watering the cultivations. Settling of wastewater in large settling tanks or ponds for a period of at least 1 month before use could ameliorate the water quality of the hazardous metal load. Further, among these three plants, the biomass produced by cocks comb (14.7 kg DW m2 year1) was the highest followed by sunflower (8.3 kg DW m2 year1) and marigold (4.1 kg DW m2 year1), and hence, the cocks comb would be the most preferred one for such phyto-amelioration process. Hence, for the purpose of phytoremediation, the option might be to use such high biomass producing plants that were also useful for economy through floriculture in the East Calcutta Wetlands.
Acknowledgments Authors thankfully acknowledge UGC DAE CSR, Kolkata Center, India for financial support. Authors also express gratitude to the Director of Technical Education and the Director of Public Instructions, Government of West Bengal, India, for cooperation and necessary support.

Conclusion The productive utilization of composite industrial wastewater for agriculture was unique to the wetland. About 60% of the total 12,500-ha wetland area was used to produce huge amount (nearly 3,70,650 kg/ha/ year) vegetables throughout the year by the use of the wastewater involving around 50,000 agro-workers in the region (Chatterjee et al. 2004, 2010). It is also true that use of such water for years heavily contaminated the farmlands and accumulation in the vegetables grown in the region was apparent. And therefore, toxic effects due to heavy metal contamination of such vegetables might also affect the food chain. However, forbidding the existing agricultural practice would immensely affect the local socioeconomic political situation of these areas. Therefore, it was felt necessary to indicate some alternative sources of earning for the community, based on the agricultural practices, as they were only used to such trade. And floriculture was thought to be the best option for such alternative agricultural practice which would reduce the metal concentration levels of contaminated lands in one hand while providing a steady source of income on the other, as flowers have a very big

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