Ecological Engineering 74 (2014) 93100Effects of organic and inorganic amendments on bio-accumulation andpartitioning of Cd in Brassica juncea and Ricinus communis

Kuldeep Bauddh a, Rana Pratap Singh b,*aCentre for Environmental Sciences, Central University of Jharkhand, Brambe, Ranchi 835205, IndiabDepartment of Environmental Science, B.B. Ambedkar University, Lucknow 226025, India

A R T I C L E I N F O

Article history:Received 1 January 2014Received in revised form 28 September 2014Accepted 6 October 2014Available online xxx

Keywords:AmendmentsCadmiumFertilizersPhytoremediationProteinProline

A B S T R A C T

The effects of different fertilizer amendments on cadmium (Cd) uptake and growth of Indian mustard(Brassica juncea L.) and castor bean (Ricinus communis L.) were investigated in this experiments. Theapplication of vermicompost, un-entrapped and entrapped forms of inorganic fertilizers (i.e., urea anddiammonium phosphate; DAP) and bio-fertilizers (Basillus subtilis and Azotobacter chrocoocum) to the soilcaused significant increase in the fresh and dry biomass of roots and shoots of both the species. Proteinand proline content in the plant leaves increased with the application of these fertilizers, however, thelevel of malondialdehyde (MDA) got decreased. Application of Cd caused decrease in protein contentwhich was found to recover with the amendments of these fertilizers. However, proline content wasfound increased with application of the fertilizers in both presence and absence of Cd in both the plants.Increased MDA content in Cd treated plants was reduced when these fertilizers were applied to the soil.Application of un-entrapped inorganic fertilizers and bio-fertilizers increased Cd uptake in the roots andshoots of both the species whereas, vermicompost and entrapped forms of these fertilizers decreased themetal accumulation. R. communis was found to be more tolerant and extracted higher amount of Cd thanthat of B. juncea. Accumulation of the metal was further increased by the application of fertilizersespecially inorganic fertilizers by R. communis.

2014 Elsevier B.V. All rights reserved.

Contents lists available at ScienceDirect

Ecological Engineering

journal homepage: www.else vie r .com/ locat e/e coleng1. Introduction

Cadmium contamination in the soil has steadily increased in lastfew decades because of mining, smelting, irrigation with industrialwastewater, electroplating, and use of chemical fertilizers andpesticides(Ghoshand Singh,2005; Zhang et al., 2010). Many physicalapproaches like electrodialysis, leaching, stabilization and landfilling, used for the remediation of heavy metals (Bayat and Sari,2010) are expensive and does not protect the ecosystem adequately(Ghosh and Singh, 2005). Phytoremediation, a plant based remedia-tion system, has emerged as an economical, eco-friendly andaesthetically acceptable technology in the recent years (Huang et al.,2011; Bauddh and Singh, 2012a,b; Santana et al., 2012; Witters et al.,2012; Olivares et al., 2013; Pandey, 2013; Kumar et al., 2014a).Phytoremediation is a natural and effective technique however, theprocess has been found to take substantial years. To address this,scientists are using chemical amendments which are found to* Corresponding author. Tel.: +91 9889121823.E-mail addresses: kuldeepenvir0811@gmail.com (K. Bauddh),

ranapsingh1@hotmail.com (R.P. Singh).

http://dx.doi.org/10.1016/j.ecoleng.2014.10.0220925-8574/ 2014 Elsevier B.V. All rights reserved.accelerate the bioaccumulation of heavy metals to some extent(Huang et al.,1997; Liphadzi et al., 2003; Garba et al., 2012a,b). Manychelants such as EDTA (ethylenediaminetetraacetic acid), CDTA(trans-1,2-diaminocyclohexane-N,N,N0,N0-tetraacetic acid), EDDHA[etylenediamine-di (o-hydroxyphenylacetic acid)], etc. have beenapplied to obtain higher efficiency in accumulating heavy metals inthe plant parts (Gr9cman et al., 2003; Meers et al., 2005; Lin et al.,2009; Wang et al., 2009; Garba et al., 2012a,b). These chemicalconstituents may alter the physicochemical and biological proper-ties of soil. The in-situ application of chelants may pose the potentialrisk of causing groundwater pollution through uncontrolled metalsolubilization and migration (Nowack 2002; Romkens et al., 2002;Madrid et al., 2003; Chen et al., 2004). It has been reported in manystudies that the application of these chemical chelants decreaseyield of plants (Blaylock et al.,1997; Chen et al., 2001). In this aspect,researchers have suggested the use of fertilizer as chelating materialto enhance the bioavailability of heavy metals for their uptake fromthe soil (Blaylock et al.,1997; Prasad and Freitas, 2003). Applicationof different inorganic and organic fertilizers in metal contaminatedsoils improve the soil fertility, physico-chemical and biologicalproperties and increase the biomass of plants which may resulthigher efficiency in extraction of the toxic metals by plants (Zaccheo

http://crossmark.dyndns.org/dialog/?doi=10.1016/j.ecoleng.2014.10.022&domain=pdfmailto:kuldeepenvir0811@gmail.commailto:ranapsingh1@hotmail.commailto:ranapsingh1@hotmail.comhttp://dx.doi.org/10.1016/j.ecoleng.2014.10.022http://dx.doi.org/10.1016/j.ecoleng.2014.10.022http://www.sciencedirect.com/science/journal/09258574www.elsevier.com/locate/ecoleng

Table 2Physicochemical properties of ex-perimental soil.

Parameters Values

pH 7.52 0.15EC(dsm1) 0.41 0.03C organic (%) 1.25 0.08N(g kg1) 1.23 0.01P(g kg1) 0.886 0.03K(g kg1) 3.01 0.19Ca(g kg1) 3.32 0.23Na(g kg1) 3.822 0.09S(mg kg1) 19.09 1.04Zn(mg kg1) 3.05 0.30Fe(mg kg1) 122.41 8.65Mn(mg kg1) 7.66 0.14Ni(mg kg1) 0.019 0.00Cd(mg kg1) 0.021 0.00Cr(mg kg1) 0.002 0.00Pb(mg kg1) 1.52 0.06Cu(mg kg1) 4.643 0.22

94 K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100et al., 2006; Jadia and Fulekar, 2008). However, such studies are rarein terms of selecting fertilizers, plants and agro-ecosystems. In thepresent work, we present differential effect of different types offertilizers (inorganic and organic) on growth of B. juncea and R.communis in terms of their responses to Cd uptake and partitioningfrom Cd contaminated soil on amendment with various fertilizernutrients.

2. Materials and methods

2.1. Plant materials and experimental design

The seeds of Indian mustard (B. juncea L.) cv Pusa Jai Kisan andcastor (R. communis L.) cv Kalpi were obtained from an authorizedoutlet of National Seeds Corporation Limited, Govt. of India. Tenseeds of each species were sown in 30-cm-diameter earthen potsfilled with 8.0 kg garden soil having different amendments. Oneplant per pot was maintained after 5 days of seedling emergence,and irrigated regularly with tap water (3 days interval with oneliter water for each pot). The experiment was conducted in sixreplicates (n = 6) to minimize the errors. The details of amend-ments (treatments) used for the experiment is given in Table 1. Thepots were kept in naturally illuminated net house of research fieldstation of university campus. All the measurements wereperformed at 30, 60 and 90 days after sowing (DAS). Physico-chemical characteristics of the experimental soil was analyzedbefore sowing of seeds and presented in Table 2.

2.2. Fertilizers used

Biofertilizers i.e., B. subtilis and A. chrocoocum and vermicom-post were obtained from Biotechnology Park, Lucknow, India.Recommended dose (RD) i.e., 160.0 kg ha1 and DAP, 80.0 kg ha1

urea and DAP and 600.0 g ha1 of each strain of biofertilizers wereused. Organic matrix entrapped fertilizers (customized fertilizers)i.e., organic matrix entrapped urea and DAP (OMEUD) and organicmatrix entrapped biofertilizers (OMEB) were prepared in our ownlab as per the method given by Kumar et al. (2012, 2013).Recommended dose of all the fertilizers were applied at the timeof sowing the seeds.

2.3. Plant growth analysis

The plants were removed from pots at 30, 60 and 90 days aftersowing (DAS) and dipped in a bucket filled with tap water. Theplant roots were washed to remove the adhering soil particles.Fresh weight (FW) of roots and shoots was taken by digital singlepan balance. The plant parts were then placed in an oven at 70 Ctill the weight became constant. The dried plant parts wereweighed to record the dry weight (DW) of roots and shoots.Table 1Details of experimental design.

Treatment Details

T0 Control: garden soilT1 Garden soil amended with 100 mT2 Vermicompost (recommended dT3 Vermicompost RD (T2) + Cd (T1)T4 Urea + DAP RD (160 and 80 kg haT5 Urea + DAP RD (T4) + Cd (T1)T6 Biofertilizers RD (consortium of T7 Biofertilizers RD (T6) + Cd (T1)T8 Organic matrix entrapped urea aT9 Organic matrix entrapped urea aT10 Organic matrix entrapped bioferT11 OMEB (T10) + Cd (T1)2.4. Estimation of protein, proline and malondialdehyde (MDA)

Fresh leaves were harvested from the mustard plants treatedwith various amendments. Protein, proline and malondialdehyde(MDA) contents were determined at 30, 60 and 90 DAS.

The protein content in the fresh leaves was estimated by themethod given by Lowry et al. (1951). Leaves (100 mg) of the controland treated plants were homogenized separately in 3 ml of 10%chilled trichloroacetic acid (TCA) in pestle and mortar andcentrifuged at 10,000 rpm for 10 min. The pellets were washedafter decanting the supernatants, and heated for 7 min with 3 ml of1 N NaOH (sodium hydroxide). It was then cooled and centrifugedagain at 10,000 rpm for 10 min. 0.5 ml of extracted sample wastaken in 2.5 ml of 0.5% CuSO4 (copper sulfate in 1% potassiumsodium tartarate), added to 48 ml of 5% Na2CO3 (sodiumcarbonate). 0.5 ml (1 N) of folin-phenol reagent was added after10 min. Incubation for 30-min developed a blue color complex inthe mixture. Absorbance was taken at 700 nm against a blank.Protein content was calculated by a standard curve made by bovineserum albumin (BSA).

Proline was determined according to the procedure followed bythe method given by Bates et al. (1973). Fresh leaves (500 mg) wereextracted with 3% aqueous 5-sulphosalicylic acid and centrifugedat 5000 rpm. The supernatant was used for the proline assay andmeasured at 520 nm. The proline content was expressed as mg g1

fresh weight (FW).The level of lipid peroxidation products in leaf samples is

expressed as MDA content and was determined as per Heath andPacker (1968). 200 mg fresh leaves were grounded in solutioncontaining 0.25% 2-thiobarbituric acid (TBA) in 10% trichloroaceticg Cd kg1 soilose; RD i.e., 4 t per hectare)

1 respectively)

Bacillus subtilis and Azotobacter chrocoocum 600 g ha1 each)

nd DAPnd DAP (T8) + Cd (T1)tilizer; OMEB

K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100 95acid (TCA) by using a mortar and pestle. After heating at 95 C for30 min, the mixture was quickly cooled in an ice bath andcentrifuged at 10,000 rpm for 10 min. The absorbance of thesupernatant was read at 532 nm and corrected for unspecificturbidity by subtracting the absorbance of the same at 600 nm. Theblank was 0.25% TBA in 10% TCA. The concentration of lipidperoxides together with oxidatively modified proteins of plantswere thus quantified in terms of MDA level using an extinctioncoefficient of 155 mM1 cm1 and expressed as nmol g1 freshweight of leaves (FW).

2.5. Estimation of cadmium

Cadmium content in the roots and shoots were estimated afterdigesting the samples in perchloric acidnitric acid mixture (1:3 v/v) by using atomic absorption spectrophotometer (AA 240 FS,Varian). Cd accumulation on the basis of per plant (by roots andshoots separately) was calculated as followed by the method of Aliet al. (2002) and Monni et al. (2000).

Total Cd extraction by roots=shootmg

plant

Dry biomass of roots=shootg metalaccumulated mg

g dry wt

by roots=shoot

2.6. Translocation factor (TF) and concentration index (CI) of Cd

Translocation factor (TF) is the ratio of the metal accumulated inshoots and roots by the plants and calculated by the equation givenby Mattina et al. (2003).Table 3Effects of different fertilizer amendments on the dry weight (g) of roots and shoots of

Treatments Roots

30 DAS 60 DAS 90 DAS

Brassica junceaT0 0.014 0.00e 0.092 0.00ef 0.112 T1 0.009 0.00h 0.068 0.00a 0.089 T2 0.0181 0.0c 0.112 0.01de 0.142 T3 0.011 0.00g 0.072 0.00fg 0.099 T4 0.022 0.00b 0.138 0.01bc 0.198 T5 0.014 0.00ef 0.111 0.00de 0.150 T6 0.016 0.00d 0.108 0.00de 0.072 T7 0.011 0.00g 0.069 0.00g 0.098 T8 0.024 0.00a 0.156 0.00b 0.221 T9 0.014 0.00e 0.122 0.01cd 0.163 T10 0.017 0.00d 0.118 0.02cd 0.079 T11 0.012 0.00fg 0.072 0.00fg 0.108

Ricinus communisT0 0.09 0.00d 0.862 0.00c 1.481 T1 0.062 0.00e 0.529 0.00d 1.012 T2 0.122 0.0b 1.124 0.01a 2.271 0T3 0.099 0.00d 0.981 0.00b 1.814 0T4 0.151 0.00a 1.316 0.01a 2.981 0T5 0.129 0.00b 1.102 0.00ab 2.519 0T6 0.102 0.00c 0.901 0.00b 1.627 0T7 0.077 0.00e 0.582 0.00d 1.1130.00fT8 0.128 0.00b 1.462 0.00a 3.125 0T9 0.096 0.00d 0.781 0.01c 2.817 0T10 0.109 0.00c 1.129 0.02a 1.916 0T11 0.079 0.00e 0.967 0.00b 1.228 0

Data was analyzed by one way analysis of variance (DMRT) at p < 0.05. Different alphabe(100 mg kg1 soil), T2: vermicompost recommended dose (RD), T3: vermicompost RD + CT7: biofertilizers RD + Cd, T8: entrapped inorganic fertilizers RD, T9: entrapped inorganic RD + Cd.Translocation factorTF CaerialCroot

where C aerial = amount of the metal accumulated in shoots(mg g1) and C root = amount of the metal accumulated in roots(mg g1)

The concentration index (CI) was calculated by dividing theamount of Cd accumulated in treatment plants by the amount of Cdaccumulated in control plants (Kiekens and Camerlynck, 1982).

Concentration indexCI

Concentration of metal in treated plantConcetration of metal in control plant

2.7. Statistical analysis

The data (n = 6) were analyzed statistically by one way analysisof variance (SPSS, Statistical package and MS Excel) using Duncansmultiple range tests (DMRT) to determine the significance ofdifferences among treatments at probability (p) 0.05 and 0.01.

3. Results

3.1. Physicochemical characteristics of experimental soil

The experimental soil was investigated for its physicochemicaland nutritional properties before the seed sowing (Table 2). Thesoil was slightly alkaline with pH 7.52, EC 0.41 dsm1 and organiccarbon 1.25%. It was found to be rich in Fe (122.41 ppm), Na(3.822 g kg1), Ca (3.32 g kg1), S (19.09 mg kg1), Mn (7.66 mgkg1) and Zn (3.05 mg kg1). The soil was having 1.23 g kg1N,Brassica juncea and Ricinus communis in the presence of Cd.

Shoots

30 DAS 60 DAS 90 DAS

0.03f 0.426 0.01c 1.260 0.04f 1.621 0.13c 0.02i 0.220 0.03e 0.716 0.05j 1.021 0.03f 0.00e 0.512 0.02b 1.426 0.02d 1.819 0.02b 0.01h 0. 0.01d 0.916 0.00i 1.120 0.02e 0.03b 0.601 0.00a 1.821 0.00b 1.992 0.02a 0.02d 0.302 0.00d 1.181 0.15g 1.482 0.03d 0.00k 0.501 0.00b 1.362 0.03e 1.620 0.03c 0.00h 0.291 0.02d 0.891 0.01i 1.102 0.05e 0.00a 0.602 0.02a 1.912 0.02a 2.016 0.03a 0.00c 0.303 0.00d 1.192 0.07g 1.526 0.02d 0.00j 0.510 0.02b 1.462 0.08c 1.828 0.08b 0.00g 0.291 0.00d 0.982 0.03h 1.112 0.01e

0.03e 0.978 0.01bc 3.781 0.04c 7.429 0.13c 0.02f 0.623 0.03d 2.457 0.05d 5.228 0.03e.00cd 1.129 0.02b 4.582 0.02b 9.321 0.02b.01d 0.992 0.01bc 3.812 0.00bc 7.061 0.02c.03b 1.326 0.00a 5.221 0.00a 10.168 0.02b.02c 1.133 0.00ab 4.692 0.15b 7.992 0.03c.00e 1.021 0.00bc 3.968 0.03b 9.018 0.03b

0.674 0.02d 2.485 0.01d 5.819 0.05e.00a 1.368 0.02a 5.982 0.02a 12.662 0.03a.00b 1.169 0.00ab 4.928 0.07ab 9.218 0.02b.00d 1.041 0.02bc 4.261 0.08b 10.225 0.08b.00f 0.756 0.00e 3.316 0.03c 6.002 0.01d

ts show significant differences between the treatments. Where: T0: control, T1: Cdd, T4: inorganic fertilizers RD, T5: inorganic fertilizers RD + Cd, T6: biofertilizers RD,fertilizers RD + Cd, T10: entrapped biofertilizers RD and T11: entrapped biofertilizers

96 K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 931000.886 g kg1 P and 3.01 g kg1K. Other metals i.e., Ni (0.019 mgkg1), Cu (4.643 mg kg1), Cr (0.002 mg kg1) and Pb (1.52 mg kg1)were present in trace amount.

3.2. Biomass production

The root and shoot dry biomass increased in differentmagnitude with the application of both inorganic and biofertilizersin conventional as well as organic matrix entrapped forms in theabsence and presence of Cd in both mustard and castor plants(Table 3). Un-entrapped and entrapped urea and DAP causedhigher plant growth than vermicompost and biofertilizerssignificantly (p < 0.01). Dry weight (DW) of the roots and shootswere also enhanced with the amendment of different fertilizers inboth the presence and absence of the metal (Table 3). Un-entrapped urea and DAP as well as organic matrix entrapped ureaand DAP (OMEUD) increased the roots and shoots DW significantly(p < 0.05) than the other fertilizers. The application of OMEUDincreased 97 and 24.36% in B. juncea and 111 and 70.44% in R.communis root and shoot DW respectively over control (nofertilizer and no Cd applied plants) at 90 DAS.

3.3. Protein, proline and malondialdehyde (MDA) contents in theleaves

Protein content in the fresh leaves of B. juncea and R. communisdecreased with the addition of Cd in the soil, however, proline andMDA were found to increase (Fig.1A and B). Application of differentfertilizers increased the protein content significantly (p < 0.05)which might be consequence of increase in protein biosynthesiseven in the presence of Cd. The leaf protein content in both theplants was generally higher in presence of urea and DAP, however,c

c

c

g g

g b

b

b

f

f f a

a a

f f

f

c

c

c

f

f f

b

b b

c

c

c

e

e

e

0

5

10

15

20

25

30

35

30 60 90

Prot

ein

cont

ent i

n th

e le

aves

(mg

g-1

FW

[A] T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

b

bc

b

cd

c

c

ab

b

b

b

b c

a

a b

b

bc

c

b

b

b

c

c

c

a

a

a

bc

bc

b

b

b

b

b

c

c

0

10

20

30

40

50

60

30 60 90

Prot

ein

cont

ent i

n th

e le

aves

(mg

g-1

FW

Time interval (Days)

[B] T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Fig.1. Effect of Cd in presence of different fertilizer amendments on protein contentin B. juncea [A] and R. communis [B] at 30, 60 and 90 DAS. Data was analyzed by oneway analysis of variance (DMRT) at p < 0.05. Different alphabets on the bars showedsignificant differences between the treatments. Where: T0 = control, T1 = Cd(100 mg kg1 soil), T2 = vermicompost recommended dose (RD), T3 = vermicompostRD + Cd, T4 = inorganic fertilizers RD, T5 = inorganic fertilizers RD + Cd, T6 = biofer-tilizers RD, T7 = biofertilizers RD + Cd, T8 = entrapped inorganic fertilizers RD,T9 = entrapped inorganic fertilizers RD + Cd, T10 = entrapped biofertilizers RD andT11 = entrapped biofertilizers RD + Cd.in presence of Cd the highest value of leaf protein was recorded inthe plants applied with vermicompost as sole added nutrient.

The metal contamination increased proline content in leaves ofboth the studied plants by 34 folds in the absence as well aspresence of different fertilizers (Fig. 2A and B). The significantlyhighest proline content (p < 0.05) was observed in the leaves of theplants grown in the soil amended with un-entrapped urea and DAP(UEUD) in B. juncea whereas in R. communis it was highest in theplants treated with OMEUD over control and other treatments atall three measurement period (30, 60 and 90 DAS) in both absenceand presence of Cd. The proline content was found to be higher in B.juncea, however, the rate of its production due to Cd stress wasmuch higher in R. communis when compared with the respectivecontrols.

Malondialdehyde (MDA) content was increased with theapplication of Cd in the soil; however, its production was reducedin the presence of different fertilizer amendments in both thespecies (Fig. 3A and B). The UEUD inhibited the production of MDAmaximum in case of B. juncea however, in case of R. communis un-entrapped biofertilizers (UEBF) reduced the production of MDAsignificantly which represents less lipid peroxidation.

3.4. Cadmium bioaccumulation

Application of UEUD as well as UEBF enhanced Cd uptake by B.juncea and R. communis (Table 4). However, entrapped forms ofthese fertilizers i.e., OMEUD and OMEBF reduced the metal uptake19.91 and 20.16% in B. juncea roots, 10.91 and 12.98 in R. communisroots, 15.53 and 15.06% in B. juncea shoots, 8.57 and 10.96% in R.communis shoots respectively (Table 4). UEUD increased Cd uptakeby 17.87, 17.56 and 10.55% by roots of B. juncea and 16.70, 35.26 and23.96% by roots of R. communis at 30, 60 and 90 DAS respectivelyover no fertilizers treated plants (T2). UEBF enhanced Cdaccumulation by 10.52, 5.90 and 9.46% in B. juncea roots and5.89, 10.08 and 16.52% in R. communis roots at 30, 60 and 90 DASe d

e

b

b b

g ef

e

de

c cd

e d

d

a

b

a

f d

f

c

b b

g e

d

c

ab cd

g e

f

d

a c

0

50

100

150

200

250

300

350

30 60 90

Prol

ine

cont

ent i

n th

e le

aves

(mg

g-1 F

W)

[A]

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

c de

ef b

c

d

c

cd e

a

b

c

d

d e a

b

b

c d

e a

a

a

d

c e

a

a

a

c

d

e a

a

a

0

5

10

15

20

25

30

35

30 60 90

Prol

ine

cont

ent i

n th

e le

aves

(mg

g-1

FW)

Time interval (Days)

[B]

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Fig. 2. Effect of Cd in presence of different fertilizer amendments on proline contentin B. juncea [A] and R. communis [B] at 30, 60 and 90 DAS. Data was analyzed by oneway analysis of variance (DMRT) at p < 0.05. Different alphabets on the bars showedsignificant differences between the treatments (the details of treatment are as inFig. 1).

f e

f

a

a

a

h

ef g

c

c

c

hi f

i

e

d

d

hi

e g

e

b

b

g f i

d

d

e

g f

h

b

d

c

0

5

10

15

20

25

30

35

30 60 90

MD

A c

onte

nt in

the

leav

es (n

mol

g-1

FW)

[A]

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

b

bc

c

a

a

a

bc

d

d

b

b

b

ab

c

d

a

b

b

b

bc

c

a

b

b

b

c

d

a

b

bc

b c

cd

a

bc

bc

0

2

4

6

8

10

12

30 60 90

MD

A c

onte

nt in

the

leav

es (

mol

g-1

FW)

Time interval (Days)

[B]

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Fig. 3. Effect of Cd in presence of different fertilizer amendments on MDA contentin B. juncea [A] and R. communis [B] at 30, 60 and 90 DAS. Data was analyzed by oneway analysis of variance (DMRT) at p < 0.05. Different alphabets on the bars showedsignificant differences between the treatments (the details of treatment are as inFig. 1).

K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100 97respectively than their respective controls (T2). Addition ofvermicompost caused significant decrease in the accumulationof Cd by both the plants. Similar pattern was also followed in caseTable 4Effects of different fertilizer amendments on Cd bioaccumulation in the roots and shoo

Treatments

Cd in roots (mg kg1 dry weight)

30 DAS 60 DAS 90 DAS

Brassica junceaT0 1.4135 0.02e 3.86235 0.23e 5.114 0.27eT1 862.245 21.59c 1105.00 47.36c 1312.666 15.62T2 1.367 0.06e 3.894 0.31e 4.838.62 0.32eT3 842.23 31.56d 1089.125 19.21d 1308.063 26.56T4 1.4535 0.02e 4.262 0.15e 5.7134 0.31eT5 1016.53 68.12a 1305.076 72.12a 1581.189 16.26T6 1.4215 0.02e 4.114 0.21e 5.529 0.26eT7 953.482 45.12b 1170.175 24.26b 1436.846 36.52T8 1.41708 0.02e 3.816 0.15e 5.0604 0.35eT9 846.6 19.22d 1078.00 39.78d 1051.159 28.72T10 1.31517 0.01e 3.314 0.21e 4.813 0.36eT11 741.913 11.62d 989.952 25.26d 1047.889 39.12

Ricinus communisT0 1.325 0.01f 1.853 0.03e 3.314 0.17eT1 351.512 25.29c 655.200 35.87c 1013.83 54.61T2 1.114 0.11f 1.613 0.11e 2.219 0.12eT3 311.389 14.26d 556.512 21.21d 873.120 19.56T4 1.819 0.06e 2.263 0.21e 3.921 0.32eT5 410.216 18.42a 885.212 32.44a 1256.811 38.92T6 1.412 0.10f 1.916 0.21e 3.812 0.27eT7 372.211 25.32b 721.241 45.56b 1181.31 44.62T8 1.162 0.08f 1.421 0.10e 2.326 0.15eT9 344.228 13.65c 581.216 26.78d 903.213 24.77T10 1.117 0.02fg 1.451 0.25e 2.162 0.16eT11 326.287 22.42d 564.312 31.56d 882.219 28.12

Data was analyzed by one way analysis of variance (DMRT) at p < 0.05. Different alphabe(100 mg kg1 soil), T2: vermicompost recommended dose (RD), T3: vermicompost RD + CT7: biofertilizers RD + Cd, T8: entrapped inorganic fertilizers RD, T9: entrapped inorganic RD + Cd.of the metal accumulation in the shoots by the application ofdifferent fertilizers. Total cadmium accumulation in roots andshoots of R. communis was much higher than that of B. juncea withthe application of different fertilizers on per plant basis (Fig. 4).

3.5. Concentration index (CI) and translocation factor (TF)

The concentration index for Cd decreased in roots and shoots ofB. juncea with increase in the plant age during 3090 DAS.Application of different fertilizers variably increased CI in bothroots and shoots with some exceptions (Table 5). However, in caseof R. communis, CI increased in roots and shoots with increase inage of plants up to 90 DAS. Application of different fertilizersincreased CI further over no fertilizer with different magnitude.Application of different form of fertilizers did not affecttranslocation factor (TF) significantly in R. communis however, itwas enhanced by the fertilizer amendments as well as with themetal exposure time in B. juncea (Fig. 5). TF ranged from 21.356 to67.744 and 17.783 to 80.796% in B. juncea and R. communisrespectively.

4. Discussion

The success of phytoremediation is based on biomass produc-tion, metal accumulation efficiency by the plants and bioavailabil-ity of heavy metals in the growing medium (McGrath, 1998). Twomodes for the phytoextraction of metals are currently under use:use of hyperaccumulator plants having high metal accumulatingcapacity (Brown et al., 1994; Kumar et al., 1995; Bauddh and Singh,2012a,b) and the utilization of high biomass producing plants withapplication of chemical chelants which can enhance phytoex-traction potential of the plants (Hernandez-Allica et al., 2008; Linet al., 2009; Wang et al., 2009; Garba et al., 2012a,b). However, theapplication of chemical chelants decrease the growth of plantts of B. juncea and R. communis.

Cd in shoots (mg kg1 dry weight)

30 DAS 60 DAS 90 DAS

0.302 0.02d 2.006 0.1f 3.412 0.23ec 243.799 11.92c 601.894 21.62c 811.1 44.15c

0.292 0.01d 1.831 0.02f 3.1613 0.14ed 241.654 9.15c 587.518 7.29de 789.3 23.26d

0.3419 .01d 2.045 0.92f 3.755 0.01ea 398.148 11.82a 725.828 24.28a 982.166 41.62a

0.327 0.03d 2.0118 0.1f 3.622 0.32eb 275.272 14.32b 676.778 13.59b 911.6 25.92b

0.289 0.01d 2.001 0.01f 3.317 0.18ed 236.728 21.62c 579.529 31.71e 685.1 21.62d

0.281 0.02d 1.863 0.02f 3.261 0.21ed 226.63 8.26c 495.435 21.49cd 688.9 12.65d

0.692 0.02d 1.100 0.1f 1.412 0.03ec 66.621 4.92c 141.812 11.62c 201.256 6.48c

0.512 0.01d 0.819 0.04f 1.018 0.12ed 56.621 2.05c 126.374 7.61de 188.262 19.62d

0.819 .03d 1.826 0.12f 2.018 0.05ea 89.826 5.82a 181.221 9.54a 261.345 21.62a

0.712 0.03d 1.512 0.05f 1.921 0.06eb 74.421 3.82b 180.211 11.59b 231.362 21.92b

0.581 0.01d 0.924 0.01f 1.119 0.14ed 61.216 4.62c 131.211 14.61e 192.263 12.62d

0.561 0.02d 0.907 0.02f 0.998 0.08ed 59.216 3.26c 122.274 13.62cd 179.2 16.55d

ts show significant differences between the treatments. Where: T0: control, T1: Cdd, T4: inorganic fertilizers RD, T5: inorganic fertilizers RD + Cd, T6: biofertilizers RD,fertilizers RD + Cd, T10: entrapped biofertilizers RD and T11: entrapped biofertilizers

0

500

1000

1500

2000

2500

3000

3500

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Cd

extr

actio

n by

roo

ts

g pe

r pl

ant

Treatments

[A] B. juncea R. communis

0

500

1000

1500

2000

2500

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Cd

extr

actio

n by

shoo

ts

g pe

r pl

ant

Treatments

[B]

B. juncea R. communis

Fig. 4. Total Cd extraction per plant by roots [A]and shoots [B]of B. juncea and R.communis in presence of different fertilizer amendments at 30, 60 and 90 DAS (thedetails of treatment are as in Fig. 1).

98 K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100(Chen et al., 2001; Lin et al., 2009; Garba et al., 2012a,b), which mayaffect the total uptake of heavy metals by the plants.

Application of different types of fertilizers have showeddifferential increase in growth and productivity of plants (Islamet al., 2012; Giansoldati et al., 2012). Application of biofertilizersincrease the growth and productivity of plants because it providesa healthy environment to the rhizosphere of metal accumulatingplants (Lodewyckx et al., 2002; Idris et al., 2004; Kumar et al.,2010). Biofertilizers has been reported to increase the mobility ofheavy metals (Lasat et al., 1996; McGrath et al., 1997). Vermicom-post increases the growth and productivity of the plants due to itshigher nutrient value and simultaneously it increases soilmicrobial activity and organic carbon of soil (Suthar et al., 2005;Mathivanan et al., 2012; Getnet and Raja, 2013). Soil amended withvermicompost produced better quality fruits and vegetables withTable 5Effects of different fertilizer amendments on concentration index for roots (CIR) and s

Treatments Concentration index for roots (CIR)

30 DAS 30 DAS 30 DAS

Brassica junceaT1 610.636 9.32 286.189 9.32 256.189 9.3T3 637.248 4.92 279.760 4.92 255.903 4.9T5 699.364 4.92 306.187 4.92 276.751 4.9T7 670.758 8.12 284.344 8.12 259.898 8.1T9 597.424 12.45 282.569 12.45 207.723 12T11 962.676 14.92 298.688 14.92 217.687 14

Ricinus communisT1 265.292 19.35 353.543 14.32 305.923 16T3 279.551 14.22 345.029 14.92 393.475 24T5 225.517 9.62 391.687 31.92 320.533 17.T7 263.238 16.15 376.423 11.12 309.893 13T9 296.238 11.45 409.023 22.45 388.312 19T11 292.110 14.22 389.116 16.22 408.057 22

T1: Cd (100 mg kg1 soil), T3: vermicompost RD + Cd, T5: inorganic fertilizers RD + Cd, T7: biofertilizers RD + Cd.less content of heavy metals or nitrate, than soil fertilized withmineral fertilizers (Mathivanan et al., 2012; Pathma and Sakthivel,2012). Several studies have been made with the application ofcustomized fertilizers for cultivation of different crops including B.juncea (Carlier et al., 2008; Kandil et al., 2010; Sharma et al., 2011;Kumar et al., 2012, 2013; Kumar et al., 2014b,c). Application ofdifferent fertilizers with or without Cd, different parametersstudied showed differential responses. Protein and prolinecontents were found to increase with the application of fertilizersboth in the presence and absence of Cd in the soil. Bentz et al.(1995) found that the protein nitrogen (N) contents of the leaveslinearly increased with the increase in level of nitrogen applied tothe plants. As N is a major structural component of protein,increase in N supply frequently lead to an increase in proteinconcentration (Brennan et al., 2000; Brennan and Bolland, 2007;Malhi and Gill, 2007; Robredo et al., 2011; Gleadow et al., 2009).Protein content in the leaves of B. juncea plants was found to bereduced with the application of Cd; however, the application ofdifferent fertilizers enhanced the soluble protein content in leavesof the plants.

Proline is an extensively studied molecule in the context ofplant responses to abiotic stresses (Pavlkov et al., 2008; Bauddhand Singh, 2012a,b). A marked increased of free proline content inabove ground biomass was detected in this study with the Cd stresswhich was further enhanced with the application of differentfertilizers. The significant increase of proline concentration wasobserved in plants with the application of fertilizers (Neuberget al., 2010). According to Atanasova (2008), the increase of prolineand alanine could serve as an indicator for unbalanced nitrogennutrition, which may be due to interference of the toxic metals.

The production of MDA in the fresh leaves was found todecrease with the application of fertilizers either in the presence orabsence of Cd in the soil. The decrease in MDA content indicates thedecrease in lipid peroxidation which is more pronounced with theaddition of fertilizers (Xue et al., 2001). The addition of nitrogenfertilizers could alleviate lipid peroxidation by increasing activitiesof antioxidant enzymes and reducing MDA content to sustainphotosynthetic function of leaves (Jiang et al., 2005; Zhang et al.,2007). Saneoka et al. (2004) stated that MDA concentrationdecreased with increased N application in water-stressed plants.Such reduction in MDA accumulation gave an indication that anadequate supply of nitrogen could alleviate the stress effect byreducing lipid peroxidation. Based on the above results, we foundthat the effects of various fertilizers on MDA concentrations in theleaves are related to nutrients availability in the growing medium.hoots (CIS) in B. juncea and R. communis.

Concentration index for roots (CIS)

30 DAS 60 DAS 90 DAS

2 807.636 12.15 300.168 13.08 237.750 8.152 827.960 24.12 320.776 14.89 249.676 9.152 1164.628 11.96 354.895 0.95 261.585 11.092 841.295 14.19 336.408 4.15 251.663 4.92.45 816.681 9.15 289.744 21.26 206.545 17.26.92 805.945 20.15 265.933 11.92 211.245 13.56

.32 96.273 16.32 128.850 11.28 142.736 6.25

.12 110.588 9.12 154.303 4.26 184.813 9.1592 109.678 13.46 99.245 8.25 129.507 9.09.89 104.524 9.09 119.187 11.12 120.417 9.52.45 105.363 3.15 142.003 10.56 171.779 11.25.32 105.554 6.25 134.811 8.92 179.578 11.00biofertilizers RD + Cd, T9: entrapped Inorganic fertilizers RD + Cd, and T11: entrapped

0

20

40

60

80

30 60 90

Tran

sloc

atio

n fa

ctor

(TF

%)

[A]

T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

0

20

40

60

80

100

30 60 90

Tran

sloc

atio

n fa

ctor

% (T

F)

Time interval (Days)

[B] T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11

Fig. 5. Translocation factor of Cd in presence of different fertilizer amendments at30, 60 and 90 DAS in B. juncea [A] and R. communis [B] (the details of treatment areas in Fig. 1).

K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100 99In this study, inorganic fertilizers (urea and DAP) andbiofertilizers (B. subtilis and A. chrocoocum) enhanced bioaccu-mulation of Cd in roots and shoots with different magnitude.Several workers have reported the increased nickel and boronuptake with the application of nitrogen and microbial fertilizers inmany plants (Schlegel et al., 1991; Ghaderian et al., 2000;Giansoldati et al., 2012). The plants used as metal bioremediatorspossess soil microorganisms in their native habitats, though thediversity and population may vary. The metal tolerant microbesmay also be available in the metal contaminated soil. The effects ofnative microbes on metal phytoremediation, however, have rarelybeen studied (Schlegel et al., 1991; Ghaderian et al., 2000). Whitinget al. (2001) found that the addition of a mixed inoculum ofMicrobacterium saperdae, Pseudomonas monteilii, and Enter-obacter cancerogenus to surface sterilized seeds of Thlaspicaerulescens sown in autoclaved soil increased the Zn concentra-tion in shoots 2 times compared with axenic controls; the totalaccumulation of Zn was enhanced 4 fold. Abou-Shanab et al.(2003a,b) have reported that soil microorganisms can produce ironchelators and siderophores that ensure iron availability, reduce soilpH and solubilize nickel compounds. Application of the vermicom-post in contaminated soil improves soil fertility and physicalproperties of the soil (Tang et al., 2003 Zheljazkov and Warman,2004). In this study, however, application of vermicompost did notincrease the metal bioaccumulation but rather reduced it. It couldattribute to adsorption of Cd by the vermicompost. Application ofthe cow dung based slow release fertilizer granules also reducedCd bioaccumulation by both the species which may be due toadsorption of Cd by the fertilizer granules. No study is available onthe effect of such customized fertilizers on Cd bioaccumulation bythe plants. The role of humic substances for reducing the uptake ofheavy metals was studied by Narancikova and Markovnikova(2003) who concluded that organic matter not only form strongcomplexes but also may retains heavy metals in exchangeableforms. Ibenake and Takenaka (2005) found that the microbialactivity on humic acids may produce smaller fragments which alsoform chelates with the heavy metal favoring the absorption byroots, which would be one of the reasons for the presence of lead inroots in all treatments with vermicompost.

5. Conclusions

The study suggests that application of inorganic fertilizers andbiofertilizers enhance the metal bioaccumulation which canreduce the time period required for the phytoremediation of theCd contaminated soil. Application of inorganic and biofertilizersenhanced tolerance mechanism in terms of protein, proline andMDA production. The vermicompost and organic matrix basedentrapped forms of inorganic and biofertilizers, reduce thebioaccumulation of Cd by B. juncea and R. communis. This approachmay be adopted to avoid the metal contamination in food chain inthe areas of lower to moderate levels of the metal. Total metalextracted by R. communis in the same time of cultivation was manyfold higher than B. juncea. It is possibly due to higher biomassproduction in castor than Indian mustard which could be increasedfurther by the application of these fertilizers.

References

Abou-Shanab, R.A., Delorme, T.A., Angle, J.S., Chaney, R.L., Ghanem, K., Moawad, H.,Ghozlan, H.A., 2003a. Phenotypic characterization of microbes in therhizosphere of Alyssum murale. Int. J. Phytoremed. 5, 367379.

Abou-Shanab, R.A., Angle, J.S., Delorme, T.A., Chaney, R.L., van Berkum, P., Moawad,H., Ghanem, K., Ghozlan, H.A., 2003b. Rhizobacterial effects on nickel extractionfrom soil and uptake by Alyssum murale. New Phytol. 158, 219224.

Ali, N.A., Bernal, M.P., Ater, M., 2002. Tolerance and bioaccumulation of copper inPhragmites australis and Zea mays. Plant Soil 239, 103111.

Atanasova, E., 2008. Effect of nitrogen sources on the nitrogenous forms andaccumulation of amino acid in head cabbage. Plant Soil Environ. 54, 6671.

Bates, L.S., Waldren, R.P., Teare, I.D., 1973. Rapid determination of proline for waterstress studies. Plant Soil 39, 205207.

Bauddh, K., Singh, R.P., 2012a. Cadmium tolerance and its phytoremediation by twooil yielding plants Ricinus communis (L.) and Brassica juncea (L.) from thecontaminated soil. Int. J. Phytorem. 14, 772785.

Bauddh, K., Singh, R.P., 2012b. Growth: tolerance efficiency and phytoremediationpotential of Ricinus communis (L.) and Brassica juncea (L.) in salinity and droughtaffected cadmium contaminated soil. Ecotox. Environ. Saf. 85, 1322.

Bayat, B., Sari, B., 2010. Comparative evaluation of microbial and chemical leachingprocesses for heavy metal removal from dewatered metal plating sludge. J.Hazard. Mater. 174, 763769.

Bentz, J.A., Reeves III, J., Barbosa, Francis, P., 1995. Within-plant variation in nitrogenand sugar content of poinsettia and its effects on the oviposition patternsurvival, and development of Bemisia argentifolii. Environ. Entomol. 24, 271277.

Brennan, R.F., Bolland, M.D.A., 2007. Effect of fertilizer phosphorus and nitrogen onthe concentrations of oil and protein in grain and the grain yield of canola(Brassica napus L.) grown in south-western Australia. Aust. J. Exp. Agric. 47, 984991.

Brennan, R.F., Mason, M.G., Walton, G.H., 2000. Effect of nitrogen fertilizer on theconcentrations of oil and protein in canola (Brassica napus) seed. J. Plant Nutr.23, 339348.

Brown, S.L., Chaney, R.L., Angle, J.S., Baker, A.J.M., 1994. Phytoremediation potentialof Thlaspi caerulescens and Bladder Campion for zinc contaminated andcadmium contaminated soil. J. Environ. Qual. 23, 11511157.

Carlier, E., Rover, M., Jaume, A.R., Rosas, S.B., 2008. Improvement of growth underfield conditions: of wheat inoculated with Pseudomonas chloraphis sub sp.aurentia SR1. World J. Microbiol. Biotechnol. 24, 26532658.

Chen, C.T., Chen, L.M., Lin, C.C., Kao, C.H., 2001. Regulation of proline accumulation indetached rice leaves exposed to copper stress. Plant Sci. 160, 283290.

Chen, Y.H., Li, X.D., Shen, Z.G., 2004. Leaching and uptake of heavy metals by tendifferent species of plants during an EDTA-assisted phytoextraction process.Chemosphere 57, 187196.

Garba, S.T., Osemeahon, A.S., Maina, H.M., Barminas, J.T., 2012a. Ethylenediamine-tetraacetate (EDTA)-assisted phytoremediation of heavy metal contaminatedsoil by Eleusine indica L. Gearth. J. Environ. Chem. Ecotoxicol. 4 (5), 103109.

Garba, S.T., Maina, H.M., Barminas, J.T., AbdulRahman, F.A., 2012b. Uptake,accumulation and translocation of cu cd, cr, and ni by dactyloctiniumaegyptium at different levels of applied edta. bulletin of environment.Pharmacol. Life Sci. 5, 4347.

Getnet, M., Raja, N., 2013. Impact of vermicompost on growth and development ofcabbage, Brassica oleracea Linn. and their sucking pest, Brevicoryne brassicaeLinn. (Homoptera: Aphididae). Res. J. Environ. Earth Sci. 5 (3), 104112.

Ghaderian, M.Y.S., Anthony, J.E.L., Baker, A.J.M., 2000. Seedling mortality of metalhyperaccumulator plants resulting from damping off by Pythium spp. NewPhytol. 146, 219224.

http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0005http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0005http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0005http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0010http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0010http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0010http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0015http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0015http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0020http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0020http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0025http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0025http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0030http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0030http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0030http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0035http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0035http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0035http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0040http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0040http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0040http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0045http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0045http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0045http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0045http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0050http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0050http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0050http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0050http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0055http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0055http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0055http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0060http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0060http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0060http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0065http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0065http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0065http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0070http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0070http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0075http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0075http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0075http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0080http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0080http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0080http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0085http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0085http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0085http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0085http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0090http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0090http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0090http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0095http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0095http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0095

100 K. Bauddh, R.P. Singh / Ecological Engineering 74 (2014) 93100Ghosh, M., Singh, S.P., 2005. A review on phytoremediation of heavy metals andutilization of its byproducts. Appl. Ecol. Environ. Res. 3 (1), 118.

Giansoldati, V., Tassi, E., Morelli, E., Gabellieri, E., Pedron, F., Barbafieri, M., 2012.Nitrogen fertilizer improves boron phytoextraction by Brassica juncea grown incontaminated sediments and alleviates plant stress. Chemosphere 87 (10),11191125.

Gleadow, R.M., Evans, J.R., McCaffery, S., Cavagnaro, T.R., 2009. Growth and nutritivevalue of cassava (Maribotesculenta Cranz) are reduced when grown in elevatedCO2. Plant Biol. 11, 7682.

Gr9cman, H., Velikonja-Bolta, , Vodnik, D., Kos, B., Le tan, D., 2003. EDTA enhancedheavy metal phytoextraction: metal accumulation, leaching and toxicity. PlantSoil 235, 105114.

Heath, R.L., Packer, L., 1968. Photoperoxidation in isolated chloroplasts: I. Kineticand stoichiometry of fatty acid peroxidation. Arch. Biochem. Biophys. 125, 189198.

Hernandez-Allica, J., Becerril, J.M., Garbisu, C., 2008. Assessment of the phytoex-traction potential of high biomass crop plants. Environ. Pollut. 152, 3240.

Huang, J.W., Chen, J.J., Berti, W.R., Cunningham, S.D.,1997. Phytoremediation of lead-contaminated soils: role of synthetic chelates in lead phytoextraction. Environ.Sci. Technol. 31, 800880.

Huang, H., Yu, N., Wang, L., Gupta, D.K., He, Z., Wang, K., Zhu, Z., Yan, X., Li, T., Yang, X.,2011. The phytoremediation potential of bioenergy crop Ricinus communis forDDTs and cadmium co-contaminated soil. Biores. Technol. 102, 1103411038.

Ibenake, S., Takenaka, C., 2005. Effects of dissolved organic matter on toxicityand bioavailability of copper from lettuce sprouts. Environ. Int. 31,603608.

Idris, R., Trifonova, R., Puschenreiter, M., Wenzel, W.W., Sessitsch, A., 2004. Bacterialcommunities associated with flowering plants of Ni hyperaccumulator Thlaspigoesingense. Appl. Environ. Microbiol. 70, 26672677.

Islam, M.R., Garcia, S.C., Horadagoda, A., 2012. Effects of irrigation and rates andtiming of nitrogen fertilizer on dry matter yield: proportions of plant fractionsof maize and nutritive value and in vitro gas production characteristics of wholecrop maize silage. Anim. Feed Sci. Technol. 172, 125135.

Jadia, C.D., Fulekar, M.H., 2008. Phytotoxicity and remediation of heavy metals byalfalfa (Medicago sativa) in soil-vermicompost media. Adv. Nat. Appl. Sci. 2 (3),141151.

Jiang, D., Fei-bo, W., Guo-ping, Z., 2005. Effects of cadmium on growth andphotosynthesis of tomato seedlings. J. Zhejiang Uni. Sci. 6B (10), 974980.

Kandil, E.A., Fawzi, M.I.F., Shahin, M.F.M., 2010. The effect of some slow releasenitrogen fertilizers on growth, nutrient status and fruiting of Mit Ghamr peachtrees. J. Am. Sci. 6, 195206.

Kiekens, L., Camerlynck, R., 1982. Transfer characteristics for uptake of heavy metalsby plants. Landwirtsch. Forsch. 39, 255261.

Kumar, P.B., Dushenkov, V.A.N., Motto, H., Raskin, I., 1995. Phytoextracton: the use ofplants to remove heavy metals from soils. Environ. Sci. Technol. 29, 12321238.

Kumar, V., Chandra, A., Singh, G., 2010. Efficacy of fly-ash based bio-fertilizers vsperfected chemical fertilizers in wheat (Triticum aestivum). Int. J. Sci. Technol. 2,3135.

Kumar, M., Bauddh, K., Sainger, M., Sainger, P.A., Singh, J.S., Singh, R.P., 2012. Increasein growth, productivity and nutritional status of rice (Oryza sativa L. c.v.Basmati) and enrichment in soil fertility applied with an organic matrixentrapped urea. J. Crop Sci. Biotechnol. 15 (2), 137144.

Kumar, M., Bauddh, K., Kumar, S., Sainger, M., Sainger, P.A., Singh, R.P., 2013. Increasein growth: productivity and nutritional status of wheat (Triticum aestivum L. C.v.Wh-711) and enrichment in soil fertility applied with organic matrix entrappedurea. J. Environ. Biol. 34, 19.

Kumar, N., Bauddh, K., Kumar, S., Dwivedi, N., Singh, D.P., Barman, S.C., 2014a. Heavymetal uptake by plants naturally grown on industrially contaminated soil andtheir phytoremediation potential. Ecol. Eng. 61, 491495.

Kumar, S., Bauddh, K., Barman, S.C., Singh, R.P., 2014b. Amendments of microbialbiofertilizers and organic substances reduces requirement of urea and DAP withenhanced nutrient availability and productivity of wheat (Triticum aestivum L.).Ecol. Eng. 71, 432437.

Kumar, M., Bauddh, K., Sainger, M., Sainger, P.A., Singh, R.P., 2014c. Increase ingrowth, productivity and nutritional status of wheat (Triticum aestivum L.) andenrichment in soil microbial population applied with biofertilizers entrappedwith organic matrix. J. Plant Nutr. http://dx.doi.org/10.1080/01904167.2014.957391.

Lasat, M.M., Baker, A.J.M., Kochain, L.V., 1996. Physiological characterization of rootZn2+ absorption and translocation to shoots in Zn hyperaccumulator and non-accumulator species of Thlaspi. Plant Physiol. 112, 17151722.

Lin, C., Liu, J., Liu, L., Zhu, T., Sheng, L., Wang, D., 2009. Soil amendment applicationfrequency contributes to phytoextraction of lead by sunflower at differentnutrient levels. Environ. Exp. Bot. 65 (23), 410416.

Liphadzi, M.S., Kirkham, M.B., Mankin, K.R., Paulsen, G.M., 2003. EDTA-assistedheavy metal uptake by poplar and sunflower grown at a long-term sewagesludge farm. Plant Soil 257, 171182.

Lodewyckx, C., Mergeay, M., Vangronsveld, J., Clijsters, H., Van Der Lelie, D., 2002.Isolation: characterization and identification of bacteria associated with thezinc hyperaccumulator Thlaspi caerulescens sub sp. calaminaria. Int. J. Phytorem.4, 101105.

Lowry, O.H., Sosenbrough, N.J., Farr, A.L., Randall, R.J., 1951. Protein measurementwith folin phenol reagent. J. Biol. Chem. 193, 265275.

Madrid, F., Liphadzi, M.S., Kirkham, M.B., 2003. Heavy metal displacement indictate-irrigated soil during phytoremediation. J. Hydrol. 272, 107119.Malhi, S.S., Gill, K.S., 2007. Interactive effects of N and S fertilizers on canola yieldand seed quality on S-deficient gray luvisol soils in northeastern Saskatchewan.Can. J. Plant Sci. 87, 211222.

Mathivanan, S., Chidambaram, A.L.A., Sundaramoorthy, P., Bakiyaraj, R., 2012. Effectof vermicompost on growth and yield of groundnut (Arachis hypogaea L.). Int. J.Environ. Biol. 2 (1), 711.

Mattina, M.J.I., Lannucci-Berger, W., Musante, C., White, J.C., 2003. Concurrent plantuptake of heavy metals and persistent organic pollutants from soil. Environ.Pollut. 124, 375378.

McGrath, S.M., Shen, Z.G., Zhao, F.J., 1997. Heavy metal uptake and chemical changesin the rhizosphere of Thlaspi caerulescens and Thlaspi ochroleucum grown incontaminates soils. Plant Soil 188, 153159.

McGrath, S.P., 1998. Phytoextraction for soil remediation. In: Brooks, R.R. (Ed.),Plants that Hyperaccumulate Heavy Metals. CAB International, Wallingford, UK,pp. 261288.

Meers, E., Ruttens, A., Hopgood, M.J., Samson, D., Tack, F.M.G., 2005. Comparison ofEDTA and EDDS as potential soil amendments for enhanced phytoextraction ofheavy metals. Chemosphere 58, 10111022.

Monni, S., Salemaa, M., Millar, N., 2000. The tolerance of Empetrum nigrum to copperand nickel. Environ. Pollut. 109, 221229.

Narancikova, G., Markovnikova, J., 2003. The influence of humic acid quality on thesorption and mobility of heavy metals. Plant Soil Environ. 12, 565571.

Neuberg, M., Pavlkov, D., Pavlk, M., Balk, J., 2010. The effect of different nitrogennutrition on proline and asparagine content in plant. Plant Soil Environ. 56 (7),305311.

Nowack, B., 2002. Environmental chemistry of aminopolycarboxylate chelatingagents. Environ. Sd. Technol. 36, 40094016.

Olivares, A.R., Carrillo-Gonzlez, R., Gonzlez-Chvez Ma del, C.A., 2013. Potential ofcastor bean (Ricinus communis L.) for phytoremediation of mine tailings and oilproduction. J. Environ. Manag. 15, 316323.

Pandey, V.C., 2013. Suitability of Ricinus communis L. cultivation for phytoreme-diation of fly ash disposal sites. Ecol. Eng. 57, 336341.

Pathma, J., Sakthivel, N., 2012. Microbial diversity of vermicompost bacteria thatexhibit useful agricultural traits and waste management potential. SpringerPlus 1, 26. doi:http://dx.doi.org/10.1186/2193-1801-1-26.

Pavlkov, D., Pavlk, M., Staszkov, L., Motyka, V., Szkov, J., Tlusto, P., Balk, J.,2008. Glutamate kinase as a potential biomarker of heavy metal stress in plants.Ecotox. Environ. Saf. 70, 223230.

Prasad, M.N.V., Freitas, H., 2003. Metal hyperaccumulation in plants?biodiversityprospecting for phytoremediation technology. Electron. J. Biotechnol. 6, 285321.

Robredo, A., Prez-Lpez, U., Miranda-Apodaca, J., Lacuesta, M., Mena-Petite, A.,Muoz-Rueda, A., 2011. Elevated CO2 reduces the drought effect on nitrogenmetabolism in barley plants during drought and subsequent recovery. Environ.Exp. Bot. 71, 399408.

Romkens, P., Bouwman, L., Japenga, J., Draaisma, C., 2002. Potentials of drawbacks ofchelate-enhanced phytoremediation of soils. Environ. Pollut. 116, 109121.

Saneoka, H., Moghaieb, R.E.A., Premachandra, G.S., Fujita, K., 2004. Nitrogennutrition and water stress effects on cell membrane stability and leaf waterrelations in Agrostis palustris Huds. Environ. Exp. Bot. 52, 131138.

Santana, K.B., de Almeida, A.F., Souza, V.L., Mangabeira, P.A.O., da, Silva D., Gomes, F.P., Dutruch, L., Loguercio, L.L., 2012. Physiological analyses of Genipa americanaL: reveals a tree with ability as phytostabilizer and rhizofilterer of chromiumions for phytoremediation of polluted watersheds. Environ. Exp. Bot. 80, 3542.

Schlegel, H.G., Cosson, J.P., Baker, A.M., 1991. Nickel hyperaccumulating plantsprovide a niche for nickel-resistant bacteria. Bot. Acta 104, 1825.

Sharma, P., Singh, G., Singh, R.P., 2011. Conservation tillage: optimal water andorganic nutrients supply enhanced soil microbial activity during wheat(Triticum aestivum) cultivation. Braz. J. Microbiol. 42, 531542.

Suthar, S.S., Watts, J., Sandhu, M., Rana, S., Kanwal, A., Gupta, D., Meena, M.S., 2005.Vermicomposting of kitchen waste by using Eisenia fetida (Savigny). Asian J.Microbiol. Biotech. Environ. Sci. 7, 541544.

Tang, S., Xi, L., Zheng, H., Li, 2003. Response to elevated CO of Indian Mustard andSunflower growing on copper contaminated soil. Bull. Environ. Contam. Toxicol.71, 988997.

Wang, X., Wang, Y., Mahmood, Q., 2009. The effect of EDDS addition on thephytoextraction efficiency from Pb contaminated soil by Sedum alfredii Hance. J.Hazard. Mater. 168 (1), 530535.

Whiting, S.N., De-Souza, M.P., Terry, N., 2001. Rhizosphere bacteria mobilize Zn forhyperaccumulation by Thlaspi caerulescens. Environ. Sci. Technol. 35, 31443150.

Witters, N., Mendelsohn, R.O., Slycken, S.V., Weyens, N., Schreurs, E., Meers, E., Tack,F., Carleer, R., Vangronsveld, J., 2012. Phytoremediation, a sustainableremediation technology? Conclusions from a case study. I: energy productionand carbon dioxide abatement. Biomass Bioenerg. 39, 454469.

Xue, T., Hartikaienen, H., Piironen, V., 2001. Antioxidative and growth-promotingeffect of selenium on senescing lettuce. Plant Oil 237, 5561.

Zaccheo, P., Crippa, L., Pasta, V., Di, M., 2006. Ammonium nutrition as a strategy forcadmium mobilization in the rhizosphere of sunflower. Plant Soil 283, 4356.

Zhang, F.Q., Wang, Y.S., Lou, Z.P., Dong, J.D., 2007. Effect of heavy metal stress onantioxidative enzymes and lipid peroxidation in leaves and roots of twomangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza).Chemosphere 67, 4450.

Zhang, S.R., Chen, M.Y., Li, T., Xu, X.X., Den, L.J., 2010. A newly found cadmiumaccumulator Malvasinensis Cavan. J. Hazard. Mater. 173, 705709.

Zheljazkov, V.D., Warman, P.R., 2004. Application of high Cu compost to dill andpeppermint. J. Agric. Food Chem. 52, 26152622.

http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0100http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0100http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0105http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0105http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0105http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0105http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0110http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0110http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0110http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0115http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0115http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0115http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0120http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0120http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0120http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0125http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0125http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0130http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0130http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0130http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0135http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0135http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0135http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0140http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0140http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0140http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0145http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0145http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0145http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0150http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0150http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0150http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0150http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0155http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0155http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0155http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0160http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0160http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0165http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0165http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0165http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0170http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0170http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0175http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0175http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0180http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0180http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0180http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0185http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0185http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0185http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0185http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0190http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0190http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0190http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0190http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0195http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0195http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0195http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0200http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0200http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0200http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0200http://dx.doi.org/10.1080/01904167.2014.957391http://dx.doi.org/10.1080/01904167.2014.957391http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0210http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0210http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0210http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0215http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0215http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0215http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0220http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0220http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0220http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0225http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0225http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0225http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0225http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0230http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0230http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0235http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0235http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0240http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0240http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0240http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0245http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0245http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0245http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0250http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0250http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0250http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0255http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0255http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0255http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0260http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0260http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0260http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0265http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0265http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0265http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0270http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0270http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0275http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0275http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0280http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0280http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0280http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0285http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0285http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0290http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0290http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0290http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0295http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0295http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0300http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0300http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0300http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0305http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0305http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0305http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0310http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0310http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0315http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0315http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0315http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0315http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0320http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0320http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0325http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0325http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0325http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0330http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0330http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0330http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0330http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0335http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0335http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0340http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0340http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0340http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0345http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0345http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0345http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0350http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0350http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0350http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0355http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0355http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0355http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0360http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0360http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0365http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0365http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0365http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0365http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0370http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0370http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0375http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0375http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0380http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0380http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0380http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0380http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0385http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0385http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0390http://refhub.elsevier.com/S0925-8574(14)00558-8/sbref0390

Effects of organic and inorganic amendments on bio-accumulation and partitioning of Cd in Brassica juncea and Ricinus communis1 Introduction2 Materials and methods2.1 Plant materials and experimental design2.2 Fertilizers used2.3 Plant growth analysis2.4 Estimation of protein, proline and malondialdehyde (MDA)2.5 Estimation of cadmium2.6 Translocation factor (TF) and concentration index (CI) of Cd2.7 Statistical analysis

3 Results3.1 Physicochemical characteristics of experimental soil3.2 Biomass production3.3 Protein, proline and malondialdehyde (MDA) contents in the leaves3.4 Cadmium bioaccumulation3.5 Concentration index (CI) and translocation factor (TF)

4 Discussion5 ConclusionsReferences