Impacts of adding biochar on nitrogen retention and bioavailability in

agricultural soil
Hao Zheng
a, b
, Zhenyu Wang
a,
, Xia Deng
a
, Stephen Herbert
b
, Baoshan Xing
b,

a
College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China
b
Stockbridge School of Agriculture, University of Massachusetts, Amherst, MA 01003, USA
a b s t r a c t a r t i c l e i n f o
Article history:
Received 22 August 2012
Received in revised form 11 April 2013
Accepted 16 April 2013
Available online 22 May 2013
Keywords:
Biochar
Nitrogen (N)
Leaching
Retention
Bioavailability
Interest in the use of biochar to improve soil productivity has rapidly increased. Nitrogen (N) loss, retention and
bioavailability in biochar-amended soils fertilized with NH
4
+
Nand NO
3

Nwere studied using leaching and pot

experiments. NO
3

N leaching from the soils fertilized with NH

4
+
N and NO
3

N was signicantly reduced by

biochar addition. NH
4
+
N leaching from the NO
3

N fertilized soil was also signicantly reduced by biochar,

while no signicant effect was observed for the NH
4
+
N fertilized soil. Also lower NH
4
+
N and NO
3

N were
leached from NO
3

N fertilized soil with maize (Zea mays L.) grown. Mitigation of N leaching loss following
biochar addition is mainly attributed to the increase in soil water holding capacity (WHC), NH
4
+
adsorption
and enhanced N immobilization. Biochar addition stimulated maize growth, both above and below ground.
Biochar also increased Nutilization efciency (NUE) of maize but decreased Naccumulation efciency (NAE), in-
dicating that biochar addition may improve N bioavailability in agricultural soils. Therefore, reduction of N
leaching, and increase of N retention and bioavailability in agricultural soils can potentially decrease the N fertil-
izer demand for crop growth.
2013 Elsevier B.V. All rights reserved.
1. Introduction
Nitrogen (N) is an essential element for plant growth and a key
agricultural input into soils. To maximize grain yield, farmers espe-
cially in developing countries often apply more N fertilizer than re-
quired for maximum crop growth (Cui et al., 2010). China is the
world's largest N fertilizer producer and consumer (Wang et al.,
2011a). In 2010, annual N fertilizer production and consumption
equated to 45.21 and 40.86 million tons (pure N), respectively, ac-
counting for about one third of the world's total production and con-
sumption (Center, 2011). In the past three decades, soil N losses have
become more serious due to the overuse and low efciency of N fer-
tilizer, although N fertilizer has signicantly contributed to the im-
provement of crop yields in China (Wang et al., 2011a). The N use
efciency by crops is 3035% in China's farmland (>50% in developed
countries), 4550% of N was lost (Ju and Zhang, 2003). The overuse
and low use efciency of N fertilizer in China as well as in other coun-
tries and regions (e.g., dead zone in the Gulf of Mexico) have already
caused serious environmental problems such as surface water eutro-
phication (Jin et al., 2005), emissions of greenhouse gas nitrous oxide
(N
2
O) (Kahrl et al., 2010), and soil acidication (Guo et al., 2010). Addi-
tionally, excessive N application represents an unnecessary economic
expenditure and waste of resource for Chinese farmers. It is estimated
that the amount of N fertilizer lost was 18.4 million tons in China in
2010 alone, causing a direct economic loss of up to $ 4.38 billion
(based on 45% loss, (Ju and Zhang, 2003)). Therefore, improving N fer-
tilizer use efciency and reducing N loss in soils are an urgent task.
N losses from the agricultural soils are resulted from gaseous
emission (N
2,
N
2
O, NO, NH
3
), and ammonium (NH
4
+
) and nitrate
(NO
3

) leaching (Robertson and Groffman, 2007). NO

3

leaching has
been the main N loss pathway from Chinese agro-ecosystem. High
NO
3

concentration in water supplies can cause eutrophication

(Huang et al., 2003) and health problems such as methemoglobine-
mia and diabetes (Feleke and Sakakibara, 2002). Therefore, technical
solutions have become obligatory to reduce N (NO
3

and NH
4
+
)
leaching losses from agricultural soils and to improve N fertilizer uti-
lization efciency for sustainable crop production. Biochar, applied to
soils has currently generated considerable interest for scientists and
policy makers (Atkinson et al., 2010), and may be a potential solution.
Biochar is produced from biomass under partial or complete exclu-
sion of oxygen, and is distinguished from charcoal by its use as a
soil amendment for improving soil quality and carbon sequestration
(Lehmann and Joseph, 2009). Biochar may improve soil physical prop-
erties, increase soil pHand cation exchange capacity (CEC), enhance nu-
trient retention and alter soil microbial populations, and augment crop
yields. This has provided the incentive to study the effects of biochar ap-
plication on soil N cycling (Lehmann et al., 2003; Ding et al., 2010;
Steiner et al., 2010; Taghizadeh-Toosi et al., 2012). A number of studies
have reported that adding biochar to soils may increase net nitrication
rate (Deluca et al., 2006), stimulate N immobilization (Rondon et al.,
Geoderma 206 (2013) 3239
Corresponding authors. Tel.: +1 413 545 5212.
E-mail addresses: wang0628@ouc.edu.cn (Z. Wang), bx@umass.edu (B. Xing).
0016-7061/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.geoderma.2013.04.018
Contents lists available at SciVerse ScienceDirect
Geoderma
j our nal homepage: www. el sevi er . com/ l ocat e/ geoder ma
2007), reduce N
2
O emission (Spokas et al., 2009), decrease NH
3
volatil-
ization (Steiner et al., 2010), enhance ammonium-oxidizing bacterial
abundance (Ball et al., 2010) and alter N availability for crops
(Lehmann et al., 2003; Steiner et al., 2008). Biochar addition has altered
N transformation following the application of N containing substances
such as bovine urine, swine manure, green-waste compost and bio-
solids (Clough et al., 2010; Laird et al., 2010b; Knowles et al., 2011).
However, the mechanisms underlying the N response to biochar addi-
tions are not well understood, but according to these reports, it is rea-
sonable to hypothesize that biochar may retain N and in theory retard
N leaching losses (Steiner et al., 2008; Ding et al., 2010; Laird et al.,
2010b; Knowles et al., 2011). Ding et al. (2010) reported that the appli-
cation of 0.5% biochar to the surface soil layer reduced cumulative
leaching losses of NH
4
+
N by 15.2% through NH
4
+
adsorption on the
biochar. Knowles et al. (2011) also reported that NO
3

N leaching
from biochar plus biosolids-amended soils was reduced to levels at or
below the control treatments, the mechanisms, however, are unclear.
In this study, we focused on the N loss and retention in a biochar-
amended soil. The specic objectives for this study were to 1) inves-
tigate the impacts of the biochar amendment on N leached from an
agricultural soil treated with different types of N fertilizer; 2) study
the N uptake by crops in the biochar-amended soil, and 3) evaluate
the N bioavailability in biochar-amended soil.
2. Materials and methods
2.1. Soil and biochar
A surface soil (020 cm) was collected from a cropland (120.38S,
36.29E) after potatoes were harvested in the Chenyang district of
Qingdao, Shandong province (China). This site was farmed in maize
vegetable rotation for the last 5 years. Urea and synthetic fertilizer
were two commonly used fertilizers in this area. The soil was
air-dried and ground to pass through a 2 mmsieve and thoroughly ho-
mogenized. Soil pH was determined in a 1:2.5 soil to water suspension
(Wang et al., 2013). Total organic carbon (TOC) was measured using an
oxidation method with potassium dichromate (Sciubba et al., 2012).
Total N(TN) content of the soil samples was determined using an auto-
matic azotometer (KDN-102F, QianjianLtd., Shanghai) (Bai et al., 2010).
Total phosphate (TP) content was spectrophotometrically determined
using phosphomolybdate blue (Parvage et al., 2012). NO
3

N and
NH
4
+
N contents in the soil were analyzed using spectrophotometric
methods with phenol disulfonic acid and indophenol blue reagent, re-
spectively (Bai et al., 2010). Bulk density was measured using a
100 cm
3
cylinder that was pressed into the soil (Zhang et al., 2010). Par-
ticle size analysis was conducted by the hydrometer method (Spokas
and Reicosky, 2009). The soil was classied as a silt loamand its proper-
ties are presented in Table 1.
Biochar was produced from giant reed (Arundo donax L.), a peren-
nial C
3
grass widely used in constructed wetlands for wastewater
treatment and ecological remediation in China, using a slow pyrolysis
method as described in Zheng et al. (2013). Giant reed was chosen
because it can produce large quantities of biomass with low nutrient
(in particular N) and pyrolysis at relatively high temperature pro-
vides biochar with high stability and even lesser N. Following char-
ring, the mass yield was recorded and the sample was milled to
pass a 2 mm sieve prior to further analysis. Total C, N, H, and O,
pH, surface area, ash content, zeta potential, total acidic oxygen-
containing groups, NH
4
+
N and NO
3

N content were characterized

as described by Zheng et al. (2013). The properties of the biochar
sample are presented in Table 1.
Table 1
Chemical and physical characteristics of the soil and biochar.
Soil Biochar
pH 6.02 Yield/% 33.4
TOC/g kg
1
17.6 pH 9.45
TN/g kg
1
1.33 C/% 73.4
NH
4
+
N/mg kg
1
63.2 H/% 3.01
NO
3

N/mg kg
1
NA
a
N/% 0.49
TP/g kg
1
1.08 O/% 23.1
Bulk density/g cm
3
1.13 NH
4
+
N/mg kg
1
17.4
Sand/% 29.2 NO
3

N/mg kg
1
3.01
Silt/% 57.2 Ash/% 10.0
Clay/% 13.6 Zeta potential/mV 44.91
CEC
b
/cmol() kg
1
15.9 CEC/cmol() kg
1
12.5
WHC
c
/% 33.6 WHC/% 95.3
Moisture content/% 2.19 Acid functional group/mmol g
1
1.16
Surface area/g m
2
2.84
a
NA: Below the detection limit.
b
CEC: cation exchange capacity, cmol() kg
1
.
c
WHC: water holding capacity, %.
Table 2
Description of experimental treatments for the leaching and pot experiments.
N fertilizer Replicate Biochar adding rate
a
0% 1% 2% 5%
(NH
4
)
2
SO
4
5 L
b
L L L
KNO
3
4 L, L + M /
c
/ L, L + M
No fertilizer 5 P P P P
a
Biochar was added at the rate of 0%, 1%, 2% and 5% by weight.
b
L indicates the leaching experiment without maize growing, L + M indicates
the leaching experiment with maize grown, P indicates the pot experiment.
c
/ means no treatment was set up at the biochar adding rate.
0
700
1400
2100
2800
3500
0 15 30 45 60 75
C
u
m
u
l
a
t
i
v
e

l
e
a
c
h
a
t
e

(
m
L
)
Time (d)
0 15 30 45 60 75
Time (d)
0%char
1%char
2%char
5%char
0
500
1000
1500
2000
2500
C
u
m
u
l
a
t
i
v
e

l
e
a
c
h
a
t
e

(
m
L
)
0%char
5%char
0%char+plant
5%char+plant
b
a
Fig. 1. The volume of cumulative leachate in the leaching experiments added with differ-
ent types of N fertilizers: (a) NH
4
+
Nfertilizer; (b) NO
3

Nfertilizer. The 0% char, 1% char,

2% and 5% char indicate that biochar was added at a rate of 0%, 1%, 2% and 5% (w/w) in the
soil columns, respectively. The 0% char + plant and 5% char + plant indicate that biochar
was added at a rate of 0% and 5% in the soil columns, respectively, where maize was
planted.
33 H. Zheng et al. / Geoderma 206 (2013) 3239
2.2. Soil column incubation and leaching
2.2.1. Preparation of soil columns
Free-draining soil columns were constructed with synthetic glass
tubes (length, 45 cm; tube internal diameter, 15 cm) and tted with
synthetic glass end caps at the bottoms (Fig. S1). A synthetic glass
plate with 0.5 mm holes was xed at 5 cm from the bottom of the
tube. A hole was drilled through the end caps and a drain tube (5 mm
in diameter) was attached to the bottom of each column. The columns
were packed with 4 kg air-dried soil by tamping the columns as the
soil was added. Quartz sand (300 g) was placed on the top and bottom
of the soil columns to prevent destruction of the soil surface during
watering. To prevent soil loss, a ne nylon mesh (b0.5 mm) was at-
tached to the base of the soil cores before packing. All columns were
packed to an initial bulk density of 1.13 g cm
3
as in the eld.
2.2.2. N leaching from the NH
4
+
N and NO
3

N fertilized soils
NH
4
+
N and NO
3

N fertilizers are the main N fertilizers on the

market and many crops prefer to take up NH
4
+
N and/or NO
3

N
from soils. Thus, the effect of biochar addition on N leaching from
the NH
4
+
N fertilized soil ((NH
4
)
2
SO
4
, equivalent to 600 mg N kg
1
soil) and NO
3

N fertilized soil (KNO

3
, equivalent to 600 mg N kg
1
soil) was studied, respectively (Table 2). Biochar was added at a
rate of 0%, 1%, 2% and 5% (w/w) in the NH
4
+
N fertilized soil, and 0%
and 5% (w/w) in the NO
3

N fertilized soil. The biochar was mixed

thoroughly with soil and N fertilizer before the columns were packed.
After the column was packed, 1.36 L of double-distilled water was
added using a perfusion tube over 3 days to reach the maximum
water holding capacity (WHC). After keeping for 7 days in a constant
temperature room (23 C and 6080% relative humidity), 250 mL of
double-distilled water was introduced on the top of each column
using a slow dripping technique (4 h) with the aid of an infusion
apparatus with a ow restricting needle mounted above the middle
of each column in every leaching event (Fig. S1). The column was
leached every three days in the rst four weeks, then every ve and
seven days in the following three and two weeks, respectively.
Moreover, in order to study the crop's roles in N leaching, maize
(Zea mays L.), a chief crop in northern China, was planted in half of
the columns packed with NO
3

N fertilized soil, thereafter referred

to as 0% char + plant and 5% char + plant. After the columns were
packed, a total of ve maize seeds were sown per column at a depth
of approximately 2 cm and then thinned to the best 3 following ger-
mination. Leaching commenced one week after germination. Because
of maize growth in this leaching experiment, the leaching was re-
duced to 12 times during the two month incubation.
The leachate fromeachcolumnwas collectedin250 mL polyethylene
bottles for 12 h after the start of a leaching event and the leachate vol-
ume was recorded. The leachate was ltered (0.45 m glass ber lter)
and analyzed within 24 h. The NO
3

N and NH
4
+
N contents in leachate
were determined using a spectrophotometric method, using phenol
disulfonic acid and Nessler's reagent, respectively (Major et al., 2010).
The pH of collected leachate was determined using a glass electrode pH
meter (Accumet BASIC AB15, Fisher Scientic Co., USA). At the end of
the leaching experiment, soil samples were collected at every 5 cm
depth from the top of the columns. A portion of each soil sample
was stored at 4 C for NH
4
+
N, NO
3

N and microorganism analysis,

0
140
280
420
560
0 15 30 45 60 75
N
H
4
+
-
N

c
u
m
u
l
a
t
i
v
e

l
e
a
c
h
i
n
g

(
m
g
)
N
O
3
-
-
N

c
u
m
u
l
a
t
i
v
e

l
e
a
c
h
i
n
g

(
m
g
)
N
O
3
-
-
N

c
u
m
u
l
a
t
i
v
e

l
e
a
c
h
i
n
g

(
m
g
)
Time (d)
0 15 30 45 60 75
Time (d)
0 15 30 45 60 75
Time (d)
0%char
1%char
2%char
5%char
0
50
100
150
200
250
300
0 20 40 60
Time (d)
0
2
4
6
0%char
5%char
0%char+plant
5%char+plant
0
500
1000
1500
2000
2500
0%char
5%char
0%char+plant
5%char+plant
b a
d c
0%char
1%char
2%char
5%char
N
H
4
+
-
N

c
u
m
u
l
a
t
i
v
e

l
e
a
c
h
i
n
g

(
m
g
)
Fig. 2. The effect of biochar on NH
4
+
N and NO
3

N leaching from the biochar-amended soils fertilized with NH

4
+
N and NO
3

N: (a) Cumulative NH
4
+
N leaching from the soil
fertilized with NH
4
+
N; (b) cumulative NO
3

N leaching from the soil fertilized with NH

4
+
N; (c) cumulative NH
4
+
N leaching from the soil fertilized with NO
3

N; (d) cumulative
NO
3

N leached from the soil fertilized with NO

3

N. The 0% char, 1% char, 2% and 5% char indicate that biochar was added at a rate of 0%, 1%, 2% and 5% (w/w) in the soil columns,
respectively. The 0% char + plant and 5% char + plant indicate that biochar was added at a rate of 0% and 5% in the soil columns, respectively, where maize was planted.
34 H. Zheng et al. / Geoderma 206 (2013) 3239
and the remaining was air-dried for pHand TN analysis. The soil pH, TN,
NO
3

Nand NH
4
+
Ncontents were measuredas describedin Section2.1.
Total microbial activity was determined using the uorescein diacetate
method (Wang et al., 2011b).
2.3. Pot experiments
Pot experiments were used to study N uptake by maize in the
same soil with biochar addition at the rates of 0%, 1%, 2% or 5%
(w/w). No additional N fertilizer was added into the soil. All the treat-
ments were replicated ve times. Each nursery pot (100 mm diameter
by 100 mm depth) was lled with 300 g of air-dried soil. The pots
were gravimetrically adjusted to maintain 60% of maximum WHC and
were left to settle in the greenhouse for 1 week before seed sowing.
Maize species was the same maize as used in the leaching experiment.
The seed germination percentage was above 98%. Soil moisture content
was kept at 60% of WHC. After 5 weeks, shoots and roots were sepa-
rately harvested. Soils were divided into rhizosphere and non-
rhizosphere soils by the hand shaking method (Wang et al., 2011b),
and TN, NO
3

N and NH
4
+
N contents were measured as described in
Section 2.1. The root parameters were obtained through WinRhizo
image analysis software (Epson Scanning and WinRhizo Pro. 2005).
The shoots and roots were dried at 65 C for dry biomass and TNanaly-
sis. TN content in shoots and roots was determined using the indophe-
nol blue photometric method after digestion with sulfuric acid and
hydrogen peroxide (Hofand et al., 1989).
2.4. Statistical analysis
All results were expressed as the mean values. Analysis of vari-
ance (ANOVA) was used to test all experimental data. The least sig-
nicant difference (LSD at P b 0.05) test was applied to assess the
differences using Statistical Product and Service Solutions Software
(SPSS 18.0).
3. Results and discussion
3.1. Effect of biochar on leachate volume
Biochar additions signicantly reduced (P b 0.05) the cumulative
leachate volume by 2.911.4% and 7.015.4% in the NH
4
+
N and NO
3

N
fertilized soils, respectively (Fig. 1). As more biochar was added, cumu-
lative leachate volume was decreased (r = 0.97, P b 0.01) (Fig. 1a).
This implied that biochar increased soil WHC (Laird et al., 2010a),
which is conrmed by the higher soil water content in the biochar-
amended soils (Fig. S2). The mechanisms for biochar-provided water
retention are mainly attributed to the changes in soil surface area,
bulk density, porosity, pore size distribution and aggregation of soil,
and these could alter percolation patterns, residence time and ow
paths of the soil solution (Major et al., 2009; Verheijen et al., 2010).
Atkinson et al. (2010) also suggested that if biochar contained sufcient
amounts of humic substances, soil WHC could be largely increased. The
biochar used in this study was fresh and had no humics, and pyrolytic
temperature (500 C) resultedina lowcontent of labile organic matters
0
50
100
150
200
250
0-5cm 5-10cm 10-15cm 15-20cm
N
H
4
+
-
N

(
m
g

k
g
-
1
)
N
O
3
-
-
N

(
m
g

k
g
-
1
)
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0%char
1%char
2%char
5%char
0
5
10
15
20
25
30
35
0%char
1%char
2%char
5%char
0
1
2
3
T
o
t
a
l

N

(
g

k
g
-
1
)
0%char 1%char
2%char 5%char
0.0
0.1
0.2
0.3
0.4
0.5
M
i
c
r
o
b
i
a
l

a
c
t
i
v
i
t
y

(
a
b
s

g
-
1

s
o
i
l
)
0%char 1%char
2%char 5%char
a
a a a
a
a
a
a
a
c
b
a
a
c
c
b
b
a
a
a
c
b
c
a
a
b
a
a
b
a
a
a
a
ab
b
c
a
c
c
b
a
c
b ab
a
a
c
b
a
a
a
a
a
b
b
b
a
b
a
ab
a
a
a
a
b a
d c
Fig. 3. The effect of biochar addition on soil N content and microbial activity in the soil fertilized with NH
4
+
N. (a) NH
4
+
N content; (b) NO
3

N content; (c) TN content; (d) total

microbial activity. The different letters among the different amount of biochar addition indicate signicant difference, which was analyzed by LSD test (P b 0.05) using Statistical
Product and Service Solutions Software (SPSS 18.0). The 0% char, 1% char, 2% and 5% char indicate that biochar was added at a rate of 0%, 1%, 2% and 5% (w/w) in the soil columns,
respectively. The 0% char + plant and 5% char + plant indicate that biochar was added at a rate of 0% and 5% in the soil columns, respectively, where maize was planted.
35 H. Zheng et al. / Geoderma 206 (2013) 3239
in the biochar, thus, direct contribution of organic matter in the biochar
may be limited. In the plant-treatments, total leachate volume was sig-
nicantly lower (P b 0.05) than that of non-plant grown treatments
(Fig. 1b). This was caused by the combined effects of the WHC increase
induced by biochar and the uptake of water by maize. Compared to the
0% char + plant treatment, less leachate was collected from the 5%
char + plant treatment during the rst month, while more leachate
was collected in the following leaching period (Fig. 1b) due to the
inhibition of plant growth in the 5% char + plant treatment, possibly
due to the inhibition of maize germination by biochar (Rogovska et
al., 2012), high water content induced by biochar application (Major
et al., 2009; Verheijen et al., 2010), or release of toxic components
(e.g., PAHs) from the biochar along with its aging in soil (Hale et al.,
2012).
Soil WHC increase may enhance nutritional benets, especially for
soil solution mobile elements (e.g. N, P and K). Additionally, WHC
0
3
6
9
12
S
o
i

N
H
4
+
-
N

(
m
g

k
g
-
1
)
S
o
i

N
O
3
-
-
N

(
m
g

k
g
-
1
)
0%char 5%char
0
4
8
12
16
0%char 5%char
0.0
0.5
1.0
1.5
2.0
2.5
0%char 5%char
0.0
0.4
0.8
1.2
1.6
2.0
2.4
non-rhizosphere rhizosphere
T
o
t
a
l

N

(
g

k
g
-
1
)
T
o
t
a
l

N

(
g

k
g
-
1
)
Soil
0%char 5%char
0
0.4
0.8
1.2
1.6
M
i
c
r
o
b
i
a
l

a
c
t
i
v
i
t
y

(
a
b
s

g
-
1

s
o
i
l
)
0%char 5%char
*
*
*
*
* *
*
*
*
*
*
*
b a
d c
e
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
0-5cm 5-10cm 10-15cm 15-20cm
Soil layer
Fig. 4. The effect of biochar addition on N content and microbial activity in soil fertilized with NO
3

N fertilizer. The asterisk indicates signicant difference, which was analyzed by

LSD test (P b 0.05) using Statistical Product and Service Solutions Software (SPSS 18.0). The 0% char, 1% char, 2% and 5% char indicate that biochar was added at a rate of 0%, 1%, 2%
and 5% (w/w) in the soil columns, respectively. The 0% char + plant and 5% char + plant indicate that biochar was added at a rate of 0% and 5% in the soil columns, respectively,
where maize was planted.
36 H. Zheng et al. / Geoderma 206 (2013) 3239
increase may reduce irrigation frequency or irrigation volume, which
in turn may reduce the risk of nutrient leaching loss, especially N.
3.2. Effect of biochar on N retention in soils fertilized with different
N fertilizers
Application of biochar to soils not only increased WHC, but also
affected the amount of N leached. The NH
4
+
N leaching from the
biochar-amended soils varied with the application of N fertilizer type
(Fig. 2). Biochar addition had no signicant inuence on the cumulative
amount of NH
4
+
N leached from the NH
4
+
N fertilized soil (Fig. 2a),
while it markedly decreased (P b 0.05) the NH
4
+
N leaching from the
NO
3

N fertilized soil (Fig. 2c). Cumulative amount of NH

4
+
N leached
from the NO
3

N fertilized soil was much smaller compared to the

NH
4
+
N fertilized soil, mainly due to the different content of NH
4
+
N
in the soils. Biochar could retain NH
4
+
Nin soils through acid functional
groups (e.g. carboxyl and hydroxyl) on its surface via cation exchange
(Clough and Condron, 2010; Zheng et al., 2013), but fresh biochar's cat-
ion exchange capacity can be limited and depends on the feedstocks
and conditions of manufacturing biochar (Taghizadeh-Toosi et al.,
2012). The NH
4
+
N content in the NH
4
+
N fertilized soil was in excess
of the maximum adsorption capacity of the soil and biochar, hence
the large amount of NH
4
+
N was leached from the NH
4
+
N fertilized
soil. But NH
4
+
N adsorption on the biochar may be the reason for the
reduction of NH
4
+
N leaching from the biochar-amended soil with
NO
3

N addition.
Biochar additions signicantly decreased (P b 0.05) the cumulative
amounts of NO
3

N leaching following either NH

4
+
N or NO
3

N fertil-
izer application (Fig. 2b, d). For the NH
4
+
N fertilized soil, the presence
of NO
3

N(Fig. 2b) indicated that nitrication occurred in the soils, and

that nitrication was slower during the rst 18 days (Fig. S3). With in-
creasing biochar rates, less NO
3

Nwas leached (r = 0.91, P = 0.09)

(Fig. 2b). For the NO
3

N fertilized soil, the NO

3

N leaching reached a
cumulative maximum relatively quickly (Fig. 2d). Biochar may adsorb
NO
3

from aqueous solution (Zheng et al., 2013), which may be one of

the reasons for the reduction of NO
3

N leaching.
Apart from the reduction of leachate volume and adsorption of
NH
4
+
N and NO
3

N on the biochar, biochar may affect N transforma-

tion in the soils (Ball et al., 2010; Clough and Condron, 2010), which
may be responsible for the reduction of N leaching from the biochar-
amended soils. The soil with biochar had higher TN (Figs. 3c, 4c, and
S4). This may have resulted fromthe enhancement of Nimmobilization
in the biochar-amended soils. It was conrmed by the increase of
microbial activity in both NH
4
+
Nand NO
3

Nfertilized soils by biochar

addition (Figs. 3d, 4e). This enhanced N immobilization process could
create a temporary reservoir of organic N, which would reduce the
potential for inorganic N leaching in highly leached soils (Lehmann et
al., 2003). However, nitrication affected by biochar in both soils was
not consistent. For the NH
4
+
N fertilized soil, more NH
4
+
N and less
NO
3

N were retained in the biochar-amended soil (Fig. 3ab), imply-

ing the inhibition of nitrication. For the NO
3

Nfertilized soil, the situ-

ation was exactly opposite (Fig. 4ab), and the enhancement of
nitrication may have occurred. In the plant-treatments, biochar addi-
tion did not alter NH
4
+
N and NO
3

N contents in both rhizosphere

and non-rhizosphere soils, but signicantly (P b 0.05) increased TN
content in non-rhizosphere soil (Fig. 4c). It may be concluded that the
biochar addition retained more N in soil with plants, because the
amount of rhizosphere soil is relatively small compared to the
non-rhizosphere. The total microbial activity was also increased by
biochar in the maize-treatment, suggesting that the enhancement of N
immobilization occurred. Ammonia volatilization in agricultural soils
is common and favored at alkaline pH and at high concentrations of
NH
4
+
N (Robertson and Groffman, 2007). The soil pH was signicantly
increased with biochar addition (Fig. S5) due to the alkaline minerals in
the biochars (Zheng et al., 2013), but not to the level (pH > 8) that
would result in signicant ammonia volatilization from the soils
(DeLuca et al., 2009). Therefore, ammonia volatilization in both leaching
experiments should be negligible. Moreover, in the plant-treatments,
the pH was not changed in the rhizosphere soil amended with or with-
out biochar compared to that of non-rhizosphere (Fig. S5c). This could
have resulted from the short time incubation with maize.
3.3. Effect of biochar on crop growth
Research has suggested that biochar applications to soils may in-
crease seed germination, plant growth, and crop yields (Lehmann et
al., 2003; Rondon et al., 2007; Graber et al., 2010). In pot experiment,
biochar addition stimulated the maize seedlings' growth (Fig. S6). The
total seedling biomass was signicantly higher in the biochar-amended
soil compared to that of the control (Fig. 5a). However, the biomass
of each plant components and total biomass did not increase with in-
creasing biochar rate (Fig. S7), suggesting that excessive biochar addition
may not leadto higher biomass andthe biochar rate shouldbe controlled
at an optimal level. The mechanisms responsible for the benecial effect
of biochar on plant growth have been proposed as the direct contribu-
tion of biochar through its intrinsic elemental and compositional nutri-
ents (e.g. N, P and K), and improvement of physical properties of the
soils resulting in benets for root growth and/or nutrient and water
0.0
0.5
1.0
1.5
0% 1% 2% 5%
M
a
i
z
e

b
i
o
m
a
s
s

(
g

p
o
t
-
1
)
Biochar amount
0% 1% 2% 5%
Biochar amount
leaf stem root
0
0.4
0.8
1.2
S
o
i
l

T
N

(
g

k
g
-
1
)
a
a
a
a
a
b
b
a
b
c
a
b
bc
A
B
B
B
a
ab
b
ab
b
Fig. 5. The effect of the biochar addition on maize biomass and soil TN in pot experi-
ments: (a) biomass; (b) soil TN. The different small and capital letters indicated signif-
icant difference analyzed by LSD test (P b 0.05) using Statistical Product and Service
Solutions Software (SPSS 18.0).
37 H. Zheng et al. / Geoderma 206 (2013) 3239
retention and acquisition (Atkinson et al., 2010). Lehmann et al. (2003)
attributed the increase of rice and cowpea biomass to K, P and, possibly,
Cu nutrition provided by their biochar. The biochar used in our study
contained little available N (20.4 mg kg
1
, Table 1) and the direct N
supply (equivalent to 4.1 mg) can be neglected. However, Graber et al.
(2010) reported that biochar application (15%) signicantly enhanced
the growth of pepper (Capsicum annuum L.) and tomato (Lycopersicum
esculentum Mill.) and their productivity, and they assigned the positive
effects to the shifts in microbial populations towards benecial plant
growthpromoting rhizobacteria or fungi, due to either chemical or phys-
ical attributes of the biochar. This was consistent with our microbial ac-
tivity data, being increased following biochar addition (Figs. 3d, 4e), and
may be used to interpret the improvement of maize growth in the
biochar-amended soils.
The improvement of plant growth in soil should come with the re-
sponse of plant root and rhizosphere, which are of importance to
water and nutrient uptake, carbohydrate storage and growth regulation
(Wang et al., 2001). Rhizosphere is the interactive interface between
plant and soil, and thus any change in root morphology is a key mecha-
nismfor the plants to adapt to soil changes suchas pHalteration and nu-
trient improvement (Hinsinger et al., 2003). In our study, the maize
roots grown in biochar-amended soil were obviously bigger than those
of the control (Fig. S6dg). The length, volume, surface area and tips of
maize roots were signicantly increased by 78.2128.2%, 36.658.9%,
30.267.3% and 7.842.9%, respectively (Table 3). Root development
is remarkably sensitive to variations in supply and distribution of in-
organic nutrients in the soil, and may quickly respond to the soil nu-
trient variation (Forde and Lorenzo, 2001). The improvement of root
morphology would certainly strengthen the nutrient bioavailability
of plants in soils (Wang et al., 2001).
3.4. Effect of biochar on N bioavailability
The addition of biochar can inuence N bioavailability and alter
bioavailability of other nutrients (Major et al., 2009; Steiner et al.,
2010; Taghizadeh-Toosi et al., 2012). Two parameters were used
here to evaluate the Nbioavailability, namely Naccumulation efciency
(NAE, the amount of N intake per unit root length/volume/SA/tips)
and N utilization efciency (NUE, the amount of biomass produced
by per unit N) (Wang et al., 2007). The NAE signicantly decreased
from 0.96 mg m
1
in the control soil to 0.290.42 mg m
1
in the
biochar-amended soils, while NUE showed a reverse trend (Table 3).
This suggested that less N was absorbed by the maize seedlings but
more biomass was produced using the equivalent amount of N in the
biochar-amended soil compared to that of the control, thus, biochar ad-
dition decreased N fertilizer demand for maize growth. Therefore, the
application of biochar in the soil enhanced the N bioavailability for
maize growth in our study.
The enhancement of N bioavailability and N retention in the biochar-
amendedsoils may result inpotential agronomic andenvironmental ben-
ets, such as reduction in fertilizer application, decline of the cost of food
production and mitigation of N
2
O emission. However, it is noted that
the effects of biochar on N leaching and bioavailability were based on
the short-term (b3 month) experiments in this study. The biochar
added in soils can be aged with the formation of oxygen-containing
functional groups (e.g. carboxyl and hydroxyl) (Cheng and Lehmann,
2009), carbon mineralization (Zimmerman, 2010) andadsorption of or-
ganic matters fromthe soils (Zhang et al., 2011) over time. The effects of
aged-biochar on N cycles may be different from the fresh biochar. For
example, the aged-biochar with newly-formed oxygen-containing
functional groups could adsorb more NH
4
+
than the fresh ones. There-
fore, works based on the long-term effects of biochar amendment on
the N cycle in soils are warranted.
4. Conclusion
Biochar addition reduced NH
4
+
N and NO
3

N leaching following
NO
3

N addition. Less NH
4
+
N and NO
3
N were leached from the
NO
3

N fertilized soil with maize grown. The mitigation of N leaching

losses following biochar addition was attributed to the soil WHC in-
crease, NH
4
+
adsorption and enhanced N immobilization. Biochar addi-
tion stimulated maize growth, both above and below ground. The NAE
and NUE were used to estimate the N bioavailability. Biochar addition
increased NUE of maize but decreased NAE. These data indicated that
biochar addition could reduce N leaching, increase N retention and
bioavailability in agricultural soils, and thus may decrease N fertilizer
demand for crop growth.
Acknowledgments
This study was supported by the Ocean Public Welfare Scientic Re-
search Project (2013418021), the Scientic and technological project of
Qingdao (10-3-3-21-nsh), and USDA Hatch Program (MAS 00982).
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://
dx.doi.org/10.1016/j.geoderma.2013.04.018.
References
Atkinson, C.J., Fitzgerald, J.D., Hipps, N.A., 2010. Potential mechanisms for achieving
agricultural benets from biochar application to temperate soils: a review. Plant
and Soil 337 (12), 118.
Bai, J.-H., Wang, Q., Hai-Feng, G., Rong, X., Wei, D., Bao-Shan, C., 2010. Spatial and
temporal distribution patterns of nitrogen in marsh soils from an inland alkaline
wetlanda case study of Fulaowenpao wetland, China. Acta Ecologica Sinica 30
(4), 210215.
Ball, P.N., MacKenzie, M.D., DeLuca, T.H., Holben, W.E., 2010. Wildre and charcoal en-
hance nitrication and ammonium-oxidizing bacterial abundance in dry montane
forest soils. Journal of Environmental Quality 39 (4), 12431253.
Center, C.N.C.I., 2011. Overview of China's Fertilizer Industry.
Cheng, C.-H., Lehmann, J., 2009. Ageing of black carbon along a temperature gradient.
Chemosphere 75 (8), 10211027.
Table 3
Rhizosphere parameters of plants in the pot experiment.
Biochar amount Length
m
Volume
cm
3
SA
a
cm
2
Tips RS
b
NAE
c
mg m
1
NUE
d
g mg
1
0% 4.27 0.89a
e
1.00 0.07a 83 13a 2116 544a 0.22 0.04a 0.96 0.23b 0.57 0.09a
1% 7.61 0.93b 1.37 0.08b 108 16ab 2282 598a 0.20 0.03a 0.42 0.02a 0.94 0.04b
2% 8.32 1.16b 1.52 0.19b 126 16b 2462 121a 0.25 0.01a 0.41 0.16a 1.02 0.05b
5% 9.74 1.86b 1.59 0.24b 139 21b 3023 648ab 0.25 0.05a 0.29 0.06a 1.18 0.06b
a
Surface area of root.
b
Ratio of root to shoot.
c
Accumulation efciency of nitrogen.
d
Utilization efciency of nitrogen.
e
Different small letter behind the values indicated signicant difference, which was analyzed by LSD test (P b 0.05) using Statistical Product and Service Solutions Software (SPSS
18.0).
38 H. Zheng et al. / Geoderma 206 (2013) 3239
Clough, T., Condron, L., 2010. Biochar and the nitrogen cycle: introduction. Journal of
Environmental Quality 39 (4), 12181223.
Clough, T., Bertram, J., Ray, J., Condron, L., O'Callaghan,, M., Sherlock, R., Wells, N., 2010.
Unweathered wood biochar impact on nitrous oxide emissions from a bovine-urine-
amended pasture soil. Soil Science Society of America Journal 74 (3), 852.
Cui, Z., Chen, X., Zhang, F., 2010. Current nitrogen management status and measures to
improve the intensive wheatmaize system in China. Ambio 39 (56), 376384.
Deluca, T.H., Mackenzie, M.D., Gundale, M.J., Holben, W.E., 2006. Wildre-produced
charcoal directly inuences nitrogen cycling in Ponderosa Pine forests. Soil Science
Society of America Journal 70 (2), 448453.
DeLuca, T.H., MacKenzie, M.D., Gundale, M.J., 2009. Biochar effects on soil nutrient trans-
formations. In: Lehmann, J., Joseph, S. (Eds.), Biochar for Environmental Manage-
ment: Science and Technology. : Biochar Effects on Soil Nutrient Transformations.
Earthscan, London, pp. 251270.
Ding, Y., Liu, Y., Wu, W., Shi, D., Yang, M., Zhong, Z., 2010. Evaluation of biochar effects
on nitrogen retention and leaching in multi-layered soil columns. Water, Air, and
Soil Pollution 213 (1), 4755.
Feleke, Z., Sakakibara, Y., 2002. A bio-electrochemical reactor coupled with adsorber for
the removal of nitrate and inhibitory pesticide. Water Research 36 (12), 30923102.
Forde, B., Lorenzo, H., 2001. The nutritional control of root development. Plant and Soil
232 (1), 5168.
Graber, E., Meller Harel, Y., Kolton, M., Cytryn, E., Silber, A., Rav David, D., Tsechansky, L.,
Borenshtein, M., Elad, Y., 2010. Biochar impact on development and productivity of
pepper and tomato grown in fertigated soilless media. Plant and Soil 337 (1), 481496.
Guo, J., Liu, X., Zhang, Y., Shen, J., Han, W., Zhang, W., Christie, P., Goulding, K., Vitousek,
P., Zhang, F., 2010. Signicant acidication in major Chinese croplands. Science 327
(5968), 10081010.
Hale, S.E., Lehmann, J., Rutherford, D., Zimmerman, A., Bachmann, R.T., Shitumbanuma,
V., O'Toole, A., Sundqvist, K.L., Arp, H.P.H., Cornelissen, G., 2012. Quantifying the
total and bioavailable PAHs and dioxins in biochars. Environmental Science and
Technology 46 (5), 28302838.
Hinsinger, P., Plassard, C., Tang, C.X., Jaillard, B., 2003. Origins of root-mediated pH
changes in the rhizosphere and their responses to environmental constraints: a
review. Plant and Soil 248 (12), 4359.
Hofand, E., Findenegg, G., Nelemans, J., 1989. Solubilization of rock phosphate by rape.
Plant and Soil 113 (2), 155160.
Huang, X., Huang, L., Yue, W., 2003. The characteristics of nutrients and eutrophication
in the Pearl River estuary, South China. Marine Pollution Bulletin 47 (16), 3036.
Jin, X., Xu, Q., Huang, C., 2005. Current status and future tendency of lake eutrophica-
tion in China. Science in China. Series C, Life Sciences 48 (2), 948954.
Ju, X., Zhang, F., 2003. Nitrate accumulation and its implication to environment in north
China. Ecology and Environment 12 (1), 2428.
Kahrl, F., Li, Y., Su, Y., Tennigkeit, T., Wilkes, A., Xu, J., 2010. Greenhouse gas emissions from
nitrogen fertilizer use in China. Environmental Science & Policy 13 (8), 688694.
Knowles, O.A., Robinson, B.H., Contangelo, A., Clucas, L., 2011. Biochar for the mitiga-
tion of nitrate leaching from soil amended with biosolids. Science of the Total
Environment 409 (17), 32063210.
Laird, D., Fleming, P., Davis, D., Horton, R., Wang, B., Karlen, D., 2010a. Impact of biochar
amendments on the quality of a typical Midwestern agricultural soil. Geoderma
158 (34), 443449.
Laird, D., Fleming, P., Wang, B., Horton, R., Karlen, D., 2010b. Biochar impact on nutrient
leaching from a Midwestern agricultural soil. Geoderma 158 (34), 436442.
Lehmann, J., Joseph, S., 2009. Biochar for Environmental Management: Science and
Technology. Earthscan, London.
Lehmann, J., Pereira da Silva, J., Steiner, C., Nehls, T., Zech, W., Glaser, B., 2003. Nutrient
availability and leaching in an archaeological Anthrosol and a Ferralsol of the Cen-
tral Amazon basin: fertilizer, manure and charcoal amendments. Plant and Soil 249
(2), 343357.
Major, J., Steiner, C., Downie, A., Lehmann, J., 2009. Biochar Effects on Nutrient Leaching.
Biochar for Environmental Management. Science and technology.Earthscan, London
271287.
Major, J., Rondon, M., Molina, D., Riha, S., Lehmann, J., 2010. Maize yield and nutrition
during 4 years after biochar application to a Colombian savanna oxisol. Plant and
Soil 333 (1), 117128.
Parvage, M., Uln, B., Eriksson, J., Strock, J., Kirchmann, H., 2012. Phosphorus availability
in soils amended with wheat residue char. Biology and Fertility of Soils 49 (2),
245250.
Robertson, G., Groffman, P., 2007. Nitrogen transformations. In: Paul, E.A., Clark, F.E.
(Eds.), Soil Microbiology and Biochemistry. Springer, New York, pp. 341364.
Rogovska, N., Laird, D., Cruse, R.M., Trabue, S., Heaton, E., 2012. Germination tests for
assessing biochar quality. Journal of Environmental Quality 41 (4), 10141022.
Rondon, M., Lehmann, J., Ramrez, J., Hurtado, M., 2007. Biological nitrogen xation by
common beans (Phaseolus vulgaris L.) increases with bio-char additions. Biology
and Fertility of Soils 43 (6), 699708.
Sciubba, L., Cavani, L., Marzadori, C., Ciavatta, C., 2012. Effect of biosolids from municipal
sewage sludge composted with rice husk on soil functionality. Biology and Fertility of
Soils 112.
Spokas, K., Reicosky, D., 2009. Impacts of sixteen different biochars on soil greenhouse
gas production. Annals of Environmental Sciences 3 (1), 179193.
Spokas, K.A., Koskinen, W.C., Baker, J.M., Reicosky, D.C., 2009. Impacts of woodchip
biochar additions on greenhouse gas production and sorption/degradation of two
herbicides in a Minnesota soil. Chemosphere 77 (4), 574581.
Steiner, C., Glaser, B., Teixeira, W.G., Lehmann, J., Blum, W.E.H., Zech, W., 2008. Nitro-
gen retention and plant uptake on a highly weathered central Amazonian Ferralsol
amended with compost and charcoal. Journal of Plant Nutrition and Soil Science
171 (6), 893899.
Steiner, C., Das, K., Melear, N., Lakly, D., 2010. Reducing nitrogen loss during poultry
litter composting using biochar. Journal of Environmental Quality 39, 12361242.
Taghizadeh-Toosi, A., Clough, T.J., Sherlock, R.R., Condron, L.M., 2012. Biochar adsorbed
ammonia is bioavailable. Plant and Soil 350 (12), 5769.
Verheijen, F., Jeffery, S., Bastos, A., Van der Velde, M., Diafas, I., 2010. Biochar Applica-
tion to Soils: A Critical Scientic Review of Effects on Soil Properties, Processes
and Functions. Joint Research Centre, Ispra, Italy.
Wang, Z., Gttlein, A., Bartonek, G., 2001. Effects of growing roots of Norway spruce
(Picea abies [L.] Karst.) and European beech (Fagus sylvatica L.) on rhizosphere
soil solution chemistry. Journal of Plant Nutrition and Soil Science 164 (1), 3541.
Wang, Z., Kelly, J., Kovar, J., 2007. Depletion of macro-nutrients from rhizosphere soil
solution by juvenile corn, cottonwood, and switchgrass plants. Plant and Soil 270
(1), 213221.
Wang, X., Cai, D., Hoogmoedc, W.B., Oenema, O., 2011a. Regional distribution of nitro-
gen fertilizer use and N-saving potential for improvement of food production and
nitrogen use efciency in China. Journal of the Science of Food and Agriculture 91,
20132023.
Wang, Z., Xu, Y., Zhao, J., Li, F., Gao, D., Xing, B., 2011b. Remediation of petroleum
contaminated soils through composting and rhizosphere degradation. Journal of
Hazardous Materials 190 (13), 677685.
Wang, Z., Zheng, H., Luo, Y., Deng, X., Herbert, S., Xing, B., 2013. Characterization and
inuence of biochars on nitrous oxide emission from agricultural soil. Environ-
mental Pollution 174, 289296.
Zhang, A., Cui, L., Pan, G., Li, L., Hussain, Q., Zhang, X., Zheng, J., Crowley, D., 2010. Effect
of biochar amendment on yield and methane and nitrous oxide emissions from a
rice paddy from Tai Lake plain, China. Agriculture, Ecosystems and Environment
139 (4), 469475.
Zhang, G., Zhang, Q., Sun, K., Liu, X., Zheng, W., Zhao, Y., 2011. Sorption of simazine to
corn straw biochars prepared at different pyrolytic temperatures. Environmental
Pollution 159 (10), 25942601.
Zheng, H., Wang, Z., Deng, X., Zhao, J., Luo, Y., Novak, J., Herbert, S., Xing, B., 2013. Char-
acteristics and nutrient values of biochars produced from giant reed at different
temperatures. Bioresource Technology 130, 463471.
Zimmerman, A.R., 2010. Abiotic and microbial oxidation of laboratory-produced black
carbon (biochar). Environmental Science and Technology 44 (4), 12951301.
39 H. Zheng et al. / Geoderma 206 (2013) 3239