Plant Soil (2011) 348:439–451

DOI 10.1007/s11104-011-0948-y

REGULAR ARTICLE

Biochar reduces the bioavailability and phytotoxicity

of heavy metals
Jin Hee Park & Girish Kumar Choppala &
Nanthi Sirangie Bolan & Jae Woo Chung &
Thammared Chuasavathi

Received: 16 May 2011 / Accepted: 1 August 2011 / Published online: 19 August 2011
# Springer Science+Business Media B.V. 2011

Abstract impact of chicken manure- and green waste-derived

Background and aims Biochar has attracted research
interest due to its ability to increase the soil carbon
pool and improve crop productivity. The objective of
this study was to evaluate the metal immobilizing
Responsible Editor: Johannes Lehmann.
J. H. Park : G. K. Choppala : N. S. Bolan (*) :
T. Chuasavathi
Centre for Environmental Risk Assessment
and Remediation, Building X,
University of South Australia,
University Boulevard,
Mawson Lakes, SA 5095, Australia
e-mail: Nanthi.Bolan@unisa.edu.au
J. H. Park : G. K. Choppala : N. S. Bolan : T. Chuasavathi
Cooperative Research Centre for Contamination
Assessment and Remediation of the Environment,
Adelaide, SA 5095, Australia
J. H. Park
Centre for Mined Land Rehabilitation,
University of Queensland,
St Lucia, QLD 4072, Australia
J. W. Chung
Department of Environmental Engineering,
Gyeongnam National University of Science
and Technology,
Dongjin-ro 33,
Jinju, Gyeongnam( 660-758, Republic of Korea
biochars, and their effectiveness in promoting plant
growth.
Methods The immobilization and phytoavailability of
Cd, Cu and Pb was examined using naturally
contaminated shooting range and spiked soils. Bio-
char samples prepared from chicken manure and
green waste were used as soil amendments.
Results Application of biochar significantly reduced
NH4NO3 extractable Cd, Cu and Pb concentrations
of soils, indicating the immobilization of these
metals. Chicken manure-derived biochar increased
plant dry biomass by 353 and 572% for shoot
and root, respectively with 1% of biochar addition.
This might be attributed to reduced toxicity of metals
and increased availability of nutrients such as P
and K. Both biochars significantly reduced Cd, Cu
and Pb accumulation by Indian mustard (Brassica
juncea), and the reduction increased with increasing
amount of biochar application except Cu concentra-
tion. Metal sequential fractionation data indicated
that biochar treatments substantially modified the
partitioning of Cd, Cu and Pb from the easily
exchangeable phase to less bioavailable organic bound
fraction.
Conclusions The results clearly showed that biochar
application was effective in metal immobilization,
thereby reducing the bioavailability and phytotoxicity
of heavy metals.
440
Keywords Chicken manure-derived biochar . Green
waste-derived biochar . Heavy metal . Immobilization .
Bioavailability
Introduction
The production of biochar from various biomass sources
has attracted tremendous research interest to sequester
carbon in the form of a thermally stabilized biomass
(Sohi et al. 2010). This is attributed to: firstly, the
increasing energy demands for their conversion to
biofuels; and secondly, concerns over greenhouse gas
emissions (Inyang et al. 2010; Laird et al. 2009).
Recent studies have highlighted that biochars are
effective soil amendments in that they improve the
agronomic values of soil (Asai et al. 2009; Hossain et
al. 2010; Laird et al. 2010). They also have the ability
to remediate organic and inorganic contaminants
(Uchimiya et al. 2010b; Yu et al. 2009). Applying
biochar leads to agronomic benefits such as improved
physical, chemical and biological properties of soils.
Biochar is a fine-grained and porous substance
produced by the slow pyrolysis of biomass at low to
medium temperatures (450 to 650°C) under oxygen-
limited conditions (Sohi et al. 2009). The source
materials of biochar may affect their performance in
terms of carbon sequestration and soil conditioning.
Whereas plant-derived biochars are considered to be a
soil conditioner rather than fertilizer, manure-derived
biochar can release nutrients and be used as both soil
fertilizer and conditioner (Uchimiya et al. 2010a).
Heavy metals such as Cd, Cu and Pb often coexist in
contaminated soils and their mobility and bioavailability
is of global concern (Uchimiya et al. 2010a, b). Biochars
are known to have a highly porous structure, contain
various functional groups and shown to be effective in
the adsorption of heavy metals, especially in aquatic
systems (Liu and Zhang 2009). However, there is a lack
of agreement over the influence of organic amendments
such as biochars on metal immobilization in soil
(Beesley et al. 2010a; Gell et al. 2011). Moreover, to
date the application of biochars to contaminated soil
systems has not been systematically investigated to any
great extent (Beesley and Marmiroli 2011). Therefore,
we examined the effects of: (a) chicken manure- and
green waste-derived biochars on the immobilization of
heavy metals in soil; and (b) biochars on plant growth
and metal mobility.
Plant Soil (2011) 348:439–451
Materials and methods
Soils and amendments
Both metal-spiked and naturally metal-contaminated
soils were used in this study. The soil used for spiking
was collected from an uncontaminated site in the
Adelaide Hills, South Australia, and spiked with Cd,
Cu and Pb at a concentration of 5, 160 and
1,000 mg kg−1, respectively. This is a concentration
1.6 times higher than that recommended by the
guideline on the Investigation Levels for Soil and
Groundwater (NEPC 1999). Additionally, two natu-
rally contaminated soils (Cd and Pb contaminated
shooting range soil from South Korea and a Cu
contaminated mine soil from Kapunda, South Australia)
were used for the immobilization experiment. The
properties of soils used in this experiment are
summarized in Table 1.
Chicken manure-derived biochar (CM) and green
waste-derived biochar (GW) were produced at a
treatment temperature of 550°C in a low temperature
pyrolysis plant by Pacific Pyrolysis at Somersby, New
South Wales (www.pacificpyrolysis.com) and were
ground (<250 μm). Biochars were analyzed for pH,
and nitrogen, carbon, hydrogen and sulfur composi-
tion using an elemental analyzer (EA1110, CE Instru-
ments). Cation exchange capacity (CEC) was
determined by a method described by Gillman and
Sumpter (1986). Surface area was analyzed using
surface area and pore size analyzer (Micromeritics,
Gemini V). The CM and GW were out-gassed in
vacuum at 105°C for 8 h prior to measurement of
surface area. The surface area of samples was
estimated using the BET (Brunauer–Emmett–Teller)
model. FTIR Spectrum was recorded using Perkin-
Elmer spectrum 400 FTIR with DRIFT at Flinders
University, South Australia. The amendment powders
were blended with spectroscopic grade KBr at the
ratio of 1:10 in an agate mortar. Spectra of the
samples were scanned between 4,000 and 500 cm−1 at
a resolution of 4.0 cm−1.
Metal immobilization
Incubation studies were performed with 100 g of Cd,
Cu and Pb spiked Adelaide Hills soil (AH), Cu mine
Kapunda soil (KC), and Cd and Pb contaminated
shooting range soil (SR). Soils were amended with
Plant Soil (2011) 348:439–451
Table 1 Properties of soils used
Soil pH EC (μS cm−1) OM (%)
Adelaide Hills soil 5.38 49.4 7.1
Copper mine soil 6.44 498 5.3
Shooting range soil 5.88 34.7 0.78
5 g (5% w/w) of CM and GW, and moisture content
was kept at 60% water holding capacity for each soil.
A control sample was prepared without biochar
treatment. After 14 days of incubation at 25°C,
samples were extracted with 1M NH4NO3 solution
ðNH4 NO3 metal for the control  NH4 NO3 metal for treated sampleÞ  100
Immobilized metalð%Þ ¼
NH4 NO3 metal for the control
The immobilization of metals was also monitored
during plant growth experiment using rhizon samplers
(see below).
Plant growth experiment
In the plant growth experiment, the Cd, Cu and Pb
spiked soil was employed to evaluate the effect of
biochars on the bioavailability of mixed metal con-
taminants. Plastic pots (600 mL capacity; 10 cm
diameter) were filled with 300 g of Cd, Cu and Pb
spiked soil, and amended with 1, 5 and 15% of CM or
GW. The biochar amended soils were incubated for
1 week at field capacity. Ten Indian mustard (Brassica
juncea) seeds were sown per pot. Rhizon samplers (one
per pot, Rhizosphere Research Products, Wageningen,
Netherlands) were placed horizontally at 2 cm from the
bottom of the pot. Each treatment was carried out in
triplicate. Pots were placed on plastic saucers to prevent
leachate draining from the soils, and the trial was carried
out in a temperature-controlled greenhouse environment
(25±3°C; 12 h light) with regular daily watering. The
number of seedlings was thinned to 5 per pot 4 days
after germination. Plants were grown for a period
of 5 weeks to monitor metal bioavailability. Pore
water samples were collected using the rhizon
sampler at 1, 2, 3, 4 and 5 weeks after the
establishment of the pot experiment, and analyzed
for metals using Inductively Coupled Plasma Mass
Spectroscopy (ICP-MS, Agilent).
441
Total Cd (mg kg−1) Total Cu (mg kg−1) Total Pb (mg kg−1)
5.0 160 1,000
0.74 1,805 161
7.2 81.3 346
(soil: solution = 1: 2.5) for 2 h, and metals were
analyzed by Inductively Coupled Plasma Optical
Emission Spectroscopy (ICP-OES, Agilent). The
immobilization of metals was calculated using the
following equation (Eq. 1):
ð1Þ
After harvesting plants, the shoots and roots were
separated, washed with Milli-Q water, oven dried and
their dry weights were recorded. The plant materials
were analysed for metal content using nitric acid
digestion (Zarcinas et al. 1987). The ground plant
material (0.100–0.500 g) was weighed directly into a
75 mL digestion tube with 5 mL of concentrated nitric
acid and left to cold digest in a fume cupboard
overnight. After keeping the samples overnight, the
tubes were heated using a temperature controlled
digestion block (AI Scientific Block Digestion System
AIM 500) programmed to slowly increase the tempera-
ture to 140°C until ca. 1 mL of digest remained in the
tube. The tubes were brought to room temperature prior
to dilution with Milli-Q water. The samples were mixed
thoroughly and filtered with a syringe filter, and analyzed
for metals by ICP-OES.
Microbial activity of soils
When the pot experiment was completed, moist and
air-dried samples from the pots were used for
microbial activity and chemical analysis, respectively.
Soil basal respiration and dehydrogenase activity
(DHA) were analyzed using moist samples to monitor
microbial activity (Anderson and Domsch 1990;
Casida et al. 1964; Singh and Singh 2005). For soil
basal respiration, 18 g (dry weight basis) of fresh soil
was weighted in a plastic tube (perforated at the top
for gas-exchange), inserted in Schott bottle containing
442 Plant Soil (2011) 348:439–451

20 mL of 0.05M NaOH solution and tightly closed.
The tubes without soil were prepared for the blank.
The bottles were incubated for 24 h at 22°C
(Anderson and Domsch 1990). After incubation, the
plastic tube was removed and 2 mL of 0.5M BaCl2
solution was added to the solution in Schott bottles.
After precipitation of BaCO3, two drops of phenol-
phthalein indicator solution were added, and the
solution was titrated with 0.03M HCl solution until
the color turned from red to colorless. The amount of
CO2 produced (mg CO2 kg−1 h−1) by soil respiration
was calculated based on the consumed HCl solution
for titration (Bloem et al. 2006).
For DHA analysis, 3 g (dry weight basis) of
fresh soil was weighed into 50 mL sterile
centrifuge tubes with 3 mL of 0.5% triphenylte-
trazolium chloride (TTC) solution in 0.1M tris
buffer (pH7.6–7.8) (Casida et al. 1964; Singh and
Singh 2005). After incubation for 24 h at 37°C,
10 mL of methanol was added to each sample to
extract produced triphenyl formazan (TPF) by DHA.
The sample suspension was centrifuged and the
absorbance of the samples was measured at 485 nm
against blank (sterile Milli-Q water).
these methods are summarized in Table 2. The
extraction was carried out in 50 mL polyethylene
centrifuge tubes; after each extraction step the
supernatant liquid was separated from the solid
phase by centrifugation at 4,000rpm for 15 min. It
was then decanted into polyethylene vessels and
stored at 4°C before analysis. The remaining residue
was washed with 10 ml of Milli-Q water, and the
washings were discarded after centrifugation. The
samples were analysed for Cd, Cu and Pb using
ICP-OES.
Statistical analysis
All experiments were conducted with three replicates.
The data collected were analyzed statistically using
SPSS 17 software. Duncan’s multiple range test was
used to compare the means of the treatments,
variability in the data was expressed as the standard
deviation, and a P < 0.05 was considered to be
statistically significant.
Results

Fractionation of metals Characteristics of amendments

Soil samples used for the chemical analysis were
dried and analyzed for pH and metal fractionation
using a sequential extraction technique. To mea-
sure pH, 5 g of each air-dried soil sample was
weighed into a 50 mL centrifuge tube and 25 mL
of Milli-Q water was added. Samples were shaken
in an end-over-end shaker for 1 h, and pH was
measured with a pH electrode after calibration.
Air-dried soil samples were sequentially extracted
with the method suggested by Tessier et al.
(1979). The reagents and operating conditions for
Biochars contain mainly carbon, a smaller proportion
of nitrogen and trace amounts of sulphur, and the pH
of biochars was alkaline (Table 3). Surface area of
CM was slightly higher than GW (Table 3). The
nature of functional groups in CM and GW was
identified by the DRIFT spectrophotometry (Fig. 1).
The peaks at 3,653 and 3,053 cm−1 indicate the presence
of hydroxyl groups. The band at 1,587 cm−1 corre-
sponds to the stretching vibrations of conjugated C=O
bonds in aromatic rings (Cao and Harris 2010). The
bands at 876 and 817 cm−1 are due to the contribution

Table 2 Extractants used in sequential extraction and nominal phases

Fraction Phase Reagent Shaking time

1 Easily exchangeable 1 M MgCl2 (pH 7) 1 h at 25°C

2 Bound to carbonates
3 Bound to Fe and Mn oxide
4 Bound to organic
5 Residual
1 M CH3COONa/CH3COOH (pH 5) 5 h at 25°C
0.04 M NH2OH·HCl in 25% (v/v) CH3COOH (pH 2) 6 h at 96°C
30% H2O2/0.02 M HNO3 (pH 2), followed by 2 h at 85°C/3 h at 85°C
3.2 M CH3COONH4/20%(v/v) HNO3
HNO3/HCl Microwave digestion
Plant Soil (2011) 348:439–451 443

Table 3 pH, elemental composition (w%), CEC and surface area of chicken manure-derived biochar (CM) and green waste-derived
biochar (GW)

Sample name pH Nitrogen (%) Carbon (%) Hydrogen (%) Sulfur (%) CEC BET
(cmol kg−1) (m2 g−1)

Chicken manure-derived biochar (CM) 8.8 2.10 51.7 2.26 0.14 238 7.27
Green waste-derived biochar (GW) 7.7 0.26 77.5 2.62 0.00 250 6.87

from C–H bond vibration in aromatic compounds
(Moreno-Castilla et al. 2000). The presence of the
above mentioned surface functional group activities are
responsible for the adsorption of metals.
Metal immobilization
The effect of biochars on the immobilization of metals
was examined by monitoring NH4NO3 extractable
metal concentration in soils treated with these amend-
ments. The addition of CM and GW significantly
reduced NH4NO3 extractable Cd concentration of
both spiked and naturally contaminated shooting
range soils, indicating metal immobilization (Table 4).
Chicken manure-derived biochar (CM) dramatically
reduced NH4NO3 extractable Cd and Pb concentra-
tions from 0.95 and 11.3 mg kg−1 to 0.11 (88.4%) and
0.73 (93.5%) mg kg−1, respectively. However, in the
case of Cu, when the initial NH4NO3 extractable Cu
concentration was low (spiked soil, AH), NH4NO3
extractable Cu concentration increased in CM
amended soil. When soil had a high Cu concentration
(Cu mine soil, KC), CM resulted in a significant
decrease in NH4NO3 extractable Cu concentration.
Green waste-derived biochar (GW) immobilized Cd,
Cu, and Pb by 30.3, 22.9 and 36.8%, respectively, for
spiked soil, and by 42.7, 0.901 and 72.9% for naturally
contaminated soil.

Biochar (Green waste)

95 Biochar (Chicken manure)

90

85

80
GW
75

70

65
%Reflectance

CM
60
3653.2

3053.8

55
817.7
878.8

50

45
3061.8

1264.7
1587.4

40
875.6

563.5

35

30
1439.5

1115.1

25

4000 3500 3000 2500 2000 1500 1000 500

Wavenumbers (cm-1)

Fig. 1 FT-IR spectra for chicken manure-derived biochar (CM) and green waste-derived biochar (GW)
444
Table 4 NH4NO3 extractable Cd, Cu and Pb concentrations
(mg kg−1) in spiked soil (AH), Cu contaminated mine soil
(KC), and Cd and Pb contaminated shooting range soil (SR)
(CM: chicken manure-derived biochar (5%), GW: green waste-
Soil Treatment Cd (mg kg−1)
AH Soil 0.955±0.0283 a
Soil + CM 0.106±0.00455 c
Soil + GW 0.666±0.0205 b
SR Soil 1.89±0.0568 a
Soil + CM 0.101±0.00153 c
Soil + GW 1.07±0.141 b
KC Soil
Soil + CM
Soil + GW
Plant growth and metal uptake
The addition of CM at 1% application rate increased
dry biomass by 353 and 572% for shoot and root,
respectively (Table 5). Green waste-derived biochar
(GW) at 15% application rate increased dry biomass
by 252 and 527% for shoot and root, respectively,
compared to the control. However, the biomass dry
weight did not increase when larger amounts of
biochar were applied, except 15% of GW treatment.
Both CM and GW were very effective in reducing
Cd and Pb concentrations of Indian mustard shoots
(Table 5). Chicken manure-derived biochar (CM)
significantly decreased Cd and Pb with increasing
level of application. Reduction in metal accumulation
by Indian mustard shoots was 74.7, 79.6 and 88.0%
for Cd, and 76.1, 82.2 and 96.3% for Pb at 1, 5 and
15% biochar addition, respectively. Green waste-
derived biochar (GW) was the most effective in
immobilizing metals when 15% of biochar was added
to the soil, achieving 67.2 and 81.6%, reduction in Cd
and Pb accumulation by Indian mustard shoots,
respectively (Table 5).
Chicken manure-derived biochar (CM) significant-
ly reduced Cd, Cu and Pb concentrations in Indian
mustard roots (Table 5). Cadmium concentration in
the roots was not significantly influenced by the
amount of biochar addition. Cadmium was evenly
distributed in both shoots and roots, but Cu and Pb
mainly accumulated in roots. Copper concentration of
the roots decreased by 53.0, 67.4 and 69.1% for CM,
and 28.7, 54.0 and 65.6% for GW with 1, 5 and 15%
Plant Soil (2011) 348:439–451
derived biochar (5%)). Each value represents the mean of three
replicates ± standard deviation, and the different letters within a
column and same soil indicate a significant difference at p<
0.05 according to Duncan’s multiple range tests using SPSS 17
Cu (mg kg−1) Pb (mg kg−1)
0.550±0.0256 b 11.3±0.408 a
0.796±0.00542 a 0.727±0.0256 c
0.424±0.0136 c 7.17±0.182 b
3.07±0.739 a
0.0000269±0.000579 b
0.832±0.289 b
222±4.79 a
47.6±16.9 b
220±66.9 a
of application, respectively, compared to the control.
The effect of biochar addition on metal concentration
in roots was more pronounced in the case of Pb,
showing 60.6, 84.2 and 88.7% decrease relative to the
control with 1, 5 and 15% of CM addition, respec-
tively. Green waste-derived biochar (GW) reduced Pb
concentration in roots by 14.6, 29.1 and 63.1% with
1, 5 and 15% application, respectively (Table 5).
Biochar addition significantly increased the plant’s
nutrient status (Table 5). Chicken manure derived-
biochar (CM) increased K in shoots by 74.1 and
100% compared to the control with 5 and 15%
addition, respectively. Phosphorus uptake was more
pronounced with CM application resulting in 257,
452 and 636% increase with 1, 5 and 15% CM
addition, respectively. Green waste-derived biochar
(GW) also significantly increased P uptake by 23.4,
119 and 216% with 1, 5 and 15% addition, respec-
tively. Nutrient uptake in roots demonstrated a trend
similar to the shoot nutrient concentrations. Potassium
concentration of roots significantly increased by 386,
1,403 and 1,516%, respectively with 1, 5 and 15% of
CM addition.
Effect of biochar on microbial activity in soil
The pH of soil increased significantly with biochar
application (Table 6). Application of biochar signifi-
cantly increased soil respiration and DHA. Soil
respiration reached its highest level when 15% of CM
was added. Dehydrogenase activity increased with
increasing addition of CM, but remained constant with
Plant Soil (2011) 348:439–451 445

990±122 e
SCM1: soil + chicken manure-derived biochar (1%), SCM5: soil + chicken manure-derived biochar (5%), SCM15: soil + chicken manure-derived biochar (15%), SGW1: soil +

5924±1664 c 1517±113 ef 1900±123 d

2688±371 de 2384±129 d
Table 5 Shoot and root dry biomass (g), and Cd, Cu, Pb, K and P concentrations (mg kg−1) in shoots and roots of Indian mustard grown in different amendments (S: soil alone,

green waste-derived biochar (1%), SGW5: soil + green waste-derived biochar (5%), SGW15: soil + green waste-derived biochar (15%)). Each value represents the mean of three

40.0±6.97 d 56.5±3.06 b 211±19.6 cd 27.6±2.78 cd 1196±69.5 c 46606±7044 c 14806±1146 b 3885±284 cd 3715±159 b

3059±368 c
6228±304 a
367±64.2 d 72575±1795 a 23948±3630 a 9048±1645 a 6506±555 a
P concentration (mg kg−1)
increasing amounts of GW, thus indicating that GW did

replicates ± standard deviation, and the different letters within a column indicate a significant difference at p<0.05 according to Duncan’s multiple range tests using SPSS 17
not affect soil microbial activity (Table 6). However,

Root
CM enhanced microbial activity, and consequently this
increased soil respiration.

1230±98.6 f

63087±6988 b 22282±2809 a 6785±136 b

7200±1307 c 4395±262 c
Shoot
Bioavailability and redistribution of metals

1482±280 d
Cadmium and Pb concentrations in pore water

7363±492 c
K concentration (mg kg−1)

decreased with increasing incubation time (Fig. 2).

Chicken manure-derived biochar (CM) and GW
Root

significantly decreased Cd and Pb concentrations in

3243±35.7 a 36241±3232 d
29731±1455 d

2769±636 ab 31155±1836 d
45767±4209 c
pore water compared to the control. However, Cu
concentration was higher in biochar amended soil
than in the control.
Shoot

Sequential extraction results reflected the results of

plant metal uptake. Addition of biochars reduced
513±139 d

2299±301 b
1276±119 c
Pb concentration (mg kg−1)

easily exchangeable fraction and increased organic

bound fraction of Cd, Cu and Pb (Fig. 3). The metals
Root

(Cd and Pb) in the treatments without biochar

16.2±1.63 cd

amendments were primarily associated with the easily

5.56±3.41 d
100±22.1 b
105±15.0 b
35.9±5.46 c
150±30.7 a

exchangeable fraction, accounting for over 80% of

Shoot

total Cd. The addition of GW at the level of 1 and 5%

showed no effect on Cd distribution in soil. However,
0.388±0.0602 a 22.6±3.49 ef 27.3±7.02 e 32.7±3.91 c 201±23.0 d
0.381±0.0977 a 13.3±0.429 f 19.1±1.61 e 35.1±7.06 c 190±9.64 d
64.9±2.15 c 80.2±2.41 a 615±23.2 a
0.433±0.0361 a 28.1±5.44 de 20.4±4.19 e 34.4±6.35 c 289±23.0 c

105±7.86 a 72.1±8.75 a 283±34.6 c

85.3±6.38 b 50.7±3.16 b 439±103 b

15% of GW reduced easily exchangeable fraction

from 84.2 to 63.1%; whereas increasing the sum of
Root

Cd associated with oxide and organic bound and

Cu concentration

residual fractions from 9.39 to 25.0%.

(mg kg−1)

Copper was strongly bound to the soil among the

Shoot

studied metals since more than 80% of its total

content was accounted for in oxide bound, organic
bound and residual fractions. The addition of CM at
1, 5 and 15% levels decreased the sum of easily
Root

exchangeable and carbonates bound fractions from

Cd concentration

18.7% to 15.2, 10.5 and 6.97%, respectively. The

0.222±0.0779 b 76.1±5.15 b
0.403±0.0837 a 36.4±1.19 d
111±11.1 a

0.239±0.0984 b 58.9±5.30 c

addition of GW showed similar results by decreasing

(mg kg−1)

those fractions to 17.9, 15.4 and 11.4% with 1, 5 and

Shoot

15% biochar, respectively. The application of 15%

CM and GW increased organic bound Cu from 46.2%
0.523±0.0687 f 0.0643±0.0135 c

to 64.2 and 46.6%, respectively.

The distribution of Pb in soil was clearly affected
by the addition of biochars. The sum of the two most
Root

labile fractions (fractions 1 and 2) for Pb decreased

Treatment Dry biomass (g)

1.51±0.0757 d
2.07±0.140 bc
2.25±0.214 ab

1.26±0.165 e
1.84±0.126 c
2.37±0.114 a

from 58.8% to 49.2, 31.2 and 16.6%, and the sum of

the two most recalcitrant fractions (fractions 4 and 5)
increased from 14.5 to 17.2, 28.2 and 48.9% with 1, 5
Shoot

and 15% CM addition, respectively. The addition of

GW reduced easily exchangeable fraction from 39.5%
SGW15
SCM15
SGW1
SGW5
SCM1
SCM5

to 38.1, 32.0 and 19.0% with 1, 5 and 15% of biochar,

respectively.
S
446 Plant Soil (2011) 348:439–451

Table 6 pH and microbial activity of chicken manure- or green
wasted-derived biochar amended soils (S: soil, SCM1: soil +
chicken manure-derived biochar (1%), SCM5: soil + chicken
manure-derived biochar (5%), SCM15: soil + chicken manure-
derived biochar (15%), SGW1: soil + green waste-derived
biochar (1%), SGW5: soil + green waste-derived biochar (5%),
SGW15: soil + green waste-derived biochar (15%)). Each value
represents the mean of three replicates ± standard deviation and
the different letters within a column indicate a significant
difference at p<0.05 according to Duncan’s multiple range tests
using SPSS 17

pH Respiration (mg CO2 kg−1 h−1) DHA (mg TPF kg−1 24 h−1)

S 5.11±0.031 g 0.305±0.016 e 2.75±0.42 d

SCM1 5.94±0.070 d 0.518±0.159 e 5.77±0.16 c
SCM5 6.77±0.015 b 2.963±1.061 ab 7.86±0.04 b
SCM15 7.51±0.021 a 3.760±0.460 a 8.96±0.92 a
SGW1 5.40±0.040 f 0.624±0.560 de 2.91±0.66 d
SGW5 5.66±0.046 e 1.634±0.451 cd 2.76±0.35 d
SGW15 6.35±0.031 c 2.484±0.644 bc 2.09±0.38 d

Discussion water-extractable Cu concentration increased, and Pb

It has often been reported that biochar improves water
retention, specific surface area, cation exchange
capacity, total N, organic C and Mehlich III extract-
able P, K, Mg and Ca (Laird et al. 2010; Sánchez et al.
2009; Thies and Rillig 2009). Biochars showed high
surface area, thereby enhancing the sorption of metals
when incorporated into soils. Uchimiya et al. (2010b)
reported that surface areas of broiler litter-derived
biochar produced at 350 and 700°C was 2.3 and
10.1 m2 g−1, respectively. In the present study, the
surface area of CM and GW was 7.27 and
6.87 m2 g−1, respectively, because it was produced
at 550°C. Chicken manure-derived biochar (CM)
showed less C/N ratio than GW. Singh et al. (2010)
characterized biochars made from different feed
stocks. In general, wood biochars had higher total C,
lower ash content, lower total N, P, K, S, Ca, Mg, Al,
Na, and Cu contents, and lower potential cation
exchange capacity and exchangeable cations than the
manure-derived biochars. The large surface area, pore
structure and functional group of biochars indicated
that metal immobilization was possible.
Biochars significantly increased Cd and Pb immo-
bilization in soil. However, biochars were not very
effective in Cu immobilization, which may be
attributed to increased Cu mobility due to increasing
dissolved organic carbon with addition of biochars
(Beesley and Dickinson 2011). Similarly, Beesley et
al. (2010b) reported that hardwood-derived biochar
amendment (8.3% w/w) significantly reduced water-
extractable Cd concentration from soil. However,
concentration was not affected by the addition of
biochar.
The immobilization of metals can be achieved by
adsorption of metals on the surface of biochars.
Adsorption of cations to organic matters and clay
mineral can be grouped into two categories, non-
specific and specific adsorption. Specific adsorption
indicates the adsorption of metals in the inner layer
forming coordination bonds to surface while non-
specific adsorption refers to adsorption of metals
by simple coulombic interaction in the diffuse electric
double layer (Abd-Elfattah and Wada 1981; Bowden
et al. 1977; Bolan et al. 1999). Most alkali and
alkaline earth cations undergo non-specific adsorp-
tion, and Cd, Pb and Zn specifically interact
with organic matter (Abd-Elfattah and Wada 1981;
Namgay et al. 2010). Therefore, metal immobilization
by biochar might occur by both specific and non-
specific adsorption.
The effect of biochar on plant growth varied with
the level of application and the nature of biochar.
Increasing amount of biochar application did not
proportionally increase the dry biomass of shoots and
roots. Matovic (2010) estimated that the optimum
biochar addition in agricultural soil ranged between
1% and 5%. The data in this experiment indicate that
in terms of plant productivity a 1% biochar applica-
tion rate is considered ideal. Both CM and GW
significantly increased shoot and root biomass of
Indian mustard, which may be attributed to reduced
metal toxicity through immobilization and supply of
nutrients. Hossain et al. (2010) tested the effect of
Plant Soil (2011) 348:439–451 447

Fig. 2 Cadmium (a), Cu 500

(b) and Pb (c) concentra- a
tions (mg L−1) in soil pore S
water in different amend- 400 SCM15
SGW15
ments with time (S: soil,
SCM15: soil + chicken

Cd concentration (mg L-1)

manure-derived biochar 300
(15%), SGW15: soil +
green waste-derived biochar
200
(15%))

100

5 10 15 20 25 30 35 40
Time (days)

900

S b
800 SCM15
SGW15
700
Cu concentration (mg L )
-1

600

500

400

300

200

100
5 10 15 20 25 30 35 40
Time (days)

2000

1800 S
c
SCM15
1600 SGW15

1400
Pb concentration (mg L-1)

1200

1000

800

600

400

200

5 10 15 20 25 30 35 40

Time (days)
448 Plant Soil (2011) 348:439–451

Fig. 3 Cadmium (a), Cu 100

(b) and Pb (c) fractiona- a
tions of soils after pot ex- Fraction 5
periment with different Fraction 4
80 Fraction 3
amendments (S: soil,
Fraction 2
SCM1: soil + chicken
Fraction 1
manure-derived biochar

Cd distribution (%)
(1%), SCM5: soil + chicken 60
manure-derived biochar
(5%), SCM15: soil + chick-
en manure-derived biochar 40
(15%), SGW1: soil + green
waste-derived biochar (1%),
SGW5: soil + green waste-
derived biochar (5%), 20
SGW15: soil + green waste-
derived biochar (15%))
0
S SCM1 SCM5 SCM15 SGW1 SGW5 SGW15
Amendments

100
b

80
Cu distribution (%)

60

Fraction 5
40 Fraction 4
Fraction 3
Fraction 2
Fraction 1
20

0
S SCM1 SCM5 SCM15 SGW1 SGW5 SGW15
Amendments

100
Fraction 5 c
Fraction 4
Fraction 3
80
Fraction 2
Fraction 1
Pb distribution (%)

60

40

20

0
S SCM1 SCM5 SCM15 SGW1 SGW5 SGW15
Amendments
Plant Soil (2011) 348:439–451
wastewater sludge biochar on the production of
cherry tomato (Lycopersicon esculentum). The addi-
tion of biochar (10 Mg ha−1) increased average dry
weight of shoot production from 61.9 to 73.8 g
plant−1, and produced a 64% higher yield compared
to the control treatment. Similarly, spring onion
cultivated in soils amended with Eucalyptus spp.
wood chips-derived biochar (1%) produced higher
biomass than those cultivated in the control soils (Yu
et al. 2009). Biochars could indirectly enhance plant
growth through supplying nutrients and trace ele-
ments, and improving soil physical and biological
properties (Lehmann et al. 2006).
Both CM and GW significantly reduced Cd and Pb
concentrations of Indian mustard with increasing
amount of amendments. However, increasing the
level of biochar application had no significant effect
on Cu concentration of shoot. It is known that Cu
concentration is associated with soluble carbon from
biochar, and pH significantly influences Cu concen-
tration (Beesley et al. 2010b). Increasing amount of
biochar might increase soluble carbon, thereby result-
ing in the mobilization of Cu. This may explain why
increasing the amount of biochar addition did not, in
fact, reduce Cu concentration in plants.
Reduced metal concentration in plants resulting
from biochar application can be attributed to both the
immobilization of bioavailable metals and ‘dilution
effect’ due to increasing plant biomass. Total Cd and
Pb uptake was significantly less when 5 and 15% of
CM was treated compared to the control. Total Cd
uptake was 62.2, 57.4 and 37.2 μg for the control,
SCM5 and SCM15, respectively, while it ranged from
75.4 to 119 μg for SCM1, SGW1, SGW5 and
SGW15. Total Pb uptake by plants showed a similar
trend. However, total Cu uptake by plants treated with
CM and GW (146–207 μg) was higher than
the control (81.5 μg). The reduction in Cd and Pb
concentrations by 5% and 15% CM was more
affected by immobilization than dilution effect
because both Cd and Pb concentrations and total
uptake were less than the control. However, reduction
in Cu concentration (Table 5) could be the result of
dilution effect by increased plant growth. Green
waste-derived biochar reduced metal concentrations
in plant tissue by increasing plant growth.
Namgay et al. (2010) reported that there was a
significant decrease in Cd concentration in maize
shoot with 15 g kg−1 of activated wood biochar, which
449
can be attributed to the formation of stable metal-
organic complexes. Cao and Harris (2010) demon-
strated that dairy manure-derived biochar was rich in
mineral elements such as N, Ca, Mg, and P in
addition to C. In our study, CM indicated lower C/N
than GW, and plant dry biomass was higher in CM
treated soil than GW. Additionally, it has been shown
that biochar application has a positive liming effect
when applied to low pH soils. The application of
biochar to acidic soils increases the soil pH, and thus
improves the immobilization of heavy metals and
nutrient availability (Hossain et al. 2010; Uchimiya et
al. 2010b).
Application of biochar significantly increased soil
respiration, which might be attributed to reduced
toxicity of metals, and increase in nutrients and plant
growth, thereby enhancing microbial activity and
decomposition of soil organic matter. Smith et al.
(2010) observed an increase in CO2 production from
soils after biochar amendment from switchgrass
(Panicum virgatum) feed stock, which increased when
larger amounts of biochar were applied. They con-
firmed that biochar contributed to the CO2 flux by
showing that the ∂13C signature of the CO2 evolved
during the incubation was the same as the ∂13C
signature of the biochar. It has often been noticed that
biochar application stimulates soil microbial activity
(Steiner et al. 2008) even though the mechanism for
biochar-induced stimulation of microbial activity is
not fully understood (Warnock et al. 2007). A
combination of physical, chemical, and biological
processes appears to contribute to microbial activity
in the soil. Warnock et al. (2007) suggested several
mechanisms for biochar-induced alteration of mycor-
rhizal fungi activity in soils and plant roots including:
(1) modification of nutrient availability and soil
physicochemical parameters; (2) enhancing the activ-
ity of other soil microorganisms that influence
mycorrhizae; (3) alteration of plant-mycorrhizal fungi
signaling processes; (4) detoxification of allelochem-
icals leading to altered root colonization by mycor-
rhizal fungi; and (5) serving as a refuge for colonizing
fungi and bacteria.
Biochars significantly decreased Cd and Pb
concentrations, but increased Cu concentration in
pore water compared to the control. Beesley et al.
(2010b) also reported that Cd concentration was
significantly reduced during the 56 days with
hardwood-derived biochar, but Cu concentration in
450
pore was significantly greater than the control soil.
Increased Cu concentration in pore water might be
associated with elevated concentrations of soluble
carbon from biochar that suggests co-mobilisation
(Beesley and Dickinson 2011). The contrasting effect
of biochar on Cd and Pb compared to Cu, was related
to the difference in the distribution of these metals in
soil. Cadmium and Pb present as easily available
fractions and the latter in more stable complexes in this
soil (Beesley et al. 2010b), as noticed in the results of
sequential extraction (Fig. 3). A substantial fraction of
Cd in the contaminated soil may be available for plant
uptake, while Cu was mainly associated with organic
bound fraction. However, increasing level of CM
application caused the immobilization of Cd, decreas-
ing their concentrations in easily exchangeable frac-
tion, which is known to be toxic to microorganisms
(Clemente and Bernal 2006).
In our study, we found that biochars are effective in
the immobilization of heavy metals even though this
effectiveness can vary depending on the type of
biochars present. Possible mechanisms for the heavy
metal immobilization by biochars are: (a) the forma-
tion of metal (hydr)oxide, carbonate, or phosphate
precipitates; (b) electrostatic interactions between
metal cations, and the activated functional groups by
increasing the pH as it was shown in FT-IR spectra;
and (c) surface chemisorption between d-electrons of
metals and delocalized π-electrons of chars (Cao et al.
2009; Uchimiya et al. 2010b). Since the soil pH
increased from 5.11 to 7.51 with 15% CM applica-
tion, this pH level can result in the precipitation of Cd
as Cd(CO3) (Mousavi et al. 2010), Cu as Cu(OH)2
(Street et al. 1977) and Pb as Pb5(PO4)3Cl (Kopittke
et al. 2008). Being transition metals, Cd and Cu have
a good coordination affinity to oxygen- and nitrogen-
bearing functional groups. Therefore, adsorption may
be attributed to coordination through phenolic –OH
and –COOH and C=N groups that present on the
surface of biochar and black carbon. Higher adsorp-
tion of Cd and Cu may be due to the electronegative
O and N atoms or π-electrons of aromatic functional
groups (Bilgic and Caliskan 2001).
Conclusions
Biochar has attracted much interest in soil amendment
to make carbon sequestration possible. However, the
Plant Soil (2011) 348:439–451
results presented here suggest that biochars have the
potential to significantly affect the behavior of metals in
soil by altering their solubility, availability, transport and
spatial distribution. This study clearly has shown that
biochar application to metal contaminated soil has the
potential of in situ remediation by immobilizing metals,
thereby reducing metal availability to the plants. In
addition, biochar improves agronomic properties by
increasing nutrient availability and microbial activity.
Chicken manure-derived biochar was more effective in
both the immobilization of metals and increasing plant
growth than green waste-derived biochar. Therefore,
chicken manure-derived biochar can be used to enhance
phytostabilization of metal contaminated soils. The
application of the findings from this study needs to be
explored under field conditions.
Acknowledgments This study was supported by the Ministry
of Education, Science and Technology (MEST) and the
Ministry of Knowledge Economy (MKE), Korea through
Gyeongnam National University of Science and Technology
as a Hub University for Industrial Collaboration (HUNIC). The
authors thank Byoung Hwan Seo, Si Young Choi and Seul Ji
Lee for laboratory assistance.
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