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Article history:
Received 12 May 2014
Received in revised form
21 November 2014
Accepted 24 November 2014
Available online 9 December 2014
We investigated how the carbon quality of soil amendments based upon their carbon (C)-to-nitrogen (N)
-ratio and their degree of aromaticity inuence soil N transformations and affect N partitioning between
soils, plants and microorganisms. A better understanding of these interactions might offer the possibility
to optimize N use efciency in agriculture. We performed a randomized pot experiment with winter
wheat and compared the inuence of naturally 13C labelled soil additives in three increasing condensation degrees, i.e. corn silage, hydrochar and pyrochar, in combination with three levels of 15N labelled
NO
3 on plant growth and N allocation. Corn silage, a lignocellulose material with a wide C-to-N-ratio and
low condensation degree, which was also used as starting material for the two other amendments,
favoured microbial growth and activity while simultaneously leading to N deciency in wheat plants. In
contrast, hydrochar and pyrochar positively inuenced plant growth independent of their C-to-N-ratio
and their degree of aromaticity. After adding hydrochar, plants did not take up the added fertilizer N but
obviously used NH
4 from mineralized hydrochar to meet their N demands. After adding pyrochar, fertilizer NO
3 was used effectively by plants and fertilizer levels were still visible in the soil, while microbial
activity was low. Our results clearly demonstrate that C quality strongly affects the N partitioning in the
plantesoilemicroorganism system. Hydrochars with a low degree of condensation that are slowly
degraded by soil microorganisms might substitute N fertilizers whereas highly condensed pyrochars
decreasing the soil microbial activity might enhance the N use efciency of plants.
2014 Elsevier Ltd. All rights reserved.
Keywords:
Hydrochar
Pyrochar
Nitrogen
15
N
Fertilizer use
Mineralization
1. Introduction
Agricultural use of soils is known for depleting their soil organic
matter (SOM) content (Post and Kwon, 2000; Aka et al., 2005).
This has adverse effects on plant growth and yield since SOM fulls
a plethora of important functions in soils, among others for soil
aeration, water holding capacity and cation exchange capacity.
Various measures have therefore been applied to minimize losses
or to restore C contents by adding amendments like compost,
biochar or other organic materials.
The various amendments added to soils in order to increase
SOM contents vary in their C-to-N ratios and contain C of different
quality: While uncharred organic matter provides mainly lignocellulose material to the soil, hydrochar adds C in the form of
267
15
12.5 and 25 kg N ha1. KNO
3 containing N (10 atom%) was added in
order to be able to track the partitioning of N between plants, microorganisms and soils. The total number of independent replicates
per treatment was three.
All pots were distributed randomly on a table in the greenhouse
and watered regularly three times per week. The appropriate watering amount was dened by previously performed watering tests
with untreated soil to investigate the water holding capacity and
the adsorption quantity of the different soils. Some representative
pots were weighed to estimate the water loss and pots were relled
accordingly. During the course of the experiment, pots with plants
progressively needed more water and, consequently, the watering
amount was adjusted to keep the soil moisture constant. This
included sometimes postponed watering in soils treated with
pyrochar, as their soil moisture persisted at a high level for a longer
period of time after watering.
2.2. Measurements
2.2.1. Plants
All pots containing seeds were checked for seedlings on days 1,
2, 3, 4, and 7 after sowing. Height measurements of all emerged
plants were performed on days 9, 11, 15, 17, 21, 28, 31 and 38/39
after sowing in order to check for differences in growth rates between the treatments. On day 38 after sowing, chlorophyll uorescence was measured on three representative leaves per pot with
a portable chlorophyll uorometer PAM-2000 (Walz Mess- und
Regeltechnik, Effeltrich, Germany) connected to a HP 200LX
blingen, Germany) in order
Palmtop PC (Hewlett Packard GmbH, Bo
to get an objective measure for plant vitality. Leaves were shaded
with leaf clips 10 min prior to the measurement in order to get a
value for minimal uorescence in the dark (F0). Via the application
of the pulse amplitude modulation technique, maximum emission
of uorescence during a saturating light pulse (Fm) and variable
uorescence (Fv FmF0) were determined. The ratio Fv/Fm is an
indicator for the performance of photosystem II and is lowered
when plants experience stress (Krause and Weis, 1991; Maxwell
and Johnson, 2000; Baker, 2008; Murchie and Lawson, 2013). It
can thus serve as an indicator for plant vitality.
Aboveground plant parts were harvested on day 39 after sowing. Leaf area of three leaves per pot was measured with a leaf
areameter LAI 3000 (Licor inc., Lincoln, Nebraska). Dry weight was
determined separately for the measured leaves and the rest of the
biomass per pot after drying the samples at 70 C for 5 days. Roots
were carefully removed from the soil, washed and dried at 70 C to
determine their dry weight per pot.
The dried plant samples were milled and subsamples were
analysed for organic C and N with a Vario Max and a Vario EL
(Elementar Analysensysteme GmbH, Hanau, Germany), respectively. Further subsamples were analysed for d15N values with a
Delta (Thermo Finnigan MAT, Bremen, Germany) coupled online
to an elemental analyser EA 1100 (CE Instruments, Milano, Italy) via
a ConFlo III (Werner et al., 1999; Steinbeiss et al., 2008a; Gubsch
et al., 2011).
Calcareous soil
Sandy soil
Corn silage
Hydrochar
Pyrochar
C [%]
N [%]
C/N
Sand [%]
Silt [%]
Clay [%]
pH 0.01 M CaCl2
d13C []
d15N []
1.8
5.3
43.7
40.6
73.7
0.20
0.29
1.22
2.45
2.20
18
9
36
17
34
9.2
50.4
e
e
e
75.1
43.8
e
e
e
15.7
5.9
e
e
e
7.5
6.8
4.6
5.4
10.6
26.37
27.82
12.33
12.80
12.75
6.81
5.94
7.05
6.88
7.46
(1:2.5)
(1:2.5)
(1:8)
(1:1)
(1:8)
268
cchar
csoil 1 cchar
(1)
(2)
Cmic
Cfum Cnonfum
kEC
(3)
(4)
The lost percentage of soil organic carbon (SOClost) was calculated in relation to the initial mass of soil organic carbon (SOCinitial)
at the start of the experiment:
SOClost % 100
SOCrem
100$SOCinitial
15
(5)
N recovery rate
Recovery rates for the 15N label (15Nrec) were calculated separately for plants and soil with obs: observed values, bg: background
value and rec: recovered label applying the 4 notation as recommended for enriched samples by Brand and Coplen (2012):
clabel
4N i
15
d15 N
Rsa 1
(6)
(6.1)
Rsa d15 N 1 *i Rref
(6.2)
R 15 N=14 N
(6.3)
(7)
showed non-homogeneous variances, non-parametric tests (KruskaleWallis and Median) were performed to conrm or reject the
results of the respective analyses of variance. Differences were
considered to be signicant if a < 0.05. Graphs were designed with
SigmaPlot10.0 (Systat Software Inc.).
3. Results
We examined the inuence of soil additives in three different
qualities (corn silage, hydrochar and pyrochar), two different soil
types and three levels of KNO
3 fertilizer on the germination success
and growth of winter wheat plants.
Seven days after sowing, germination success on soils treated
with hydrochar was signicantly (p < 0.001) lower (48%) compared
to the two other treatments (75% on corn silage and 77% on pyrochar). This difference was still evident at the end of the experiment
e until day 31 after sowing, germination rate on soils treated with
hydrochar reached 74% and thus was still considerably lower than
on soils treated with corn silage (91%) or pyrochar (93%). A comparable negative effect of hydrochar on plant growth could be
detected during the rst half of the experiment. Nine days after
sowing, plants on soils treated with hydrochar only achieved an
average height of 4 cm, while plants on corn silage reached 6.6 cm
and plants on soils treated with pyrochar 7.5 cm. On days 17 and 21
after sowing, differences in height growth between corn silage and
hydrochar treatments disappeared. Thus, height growth of plants
on hydrochar signicantly surpassed that of the plants on corn
silage at the end of the experiment despite a slower growth in the
initial phase, but remained below that of plants grown on hydrochar (Table 2).
Similarly, shoot dry weight of plants grown on soils with pyrochar reached 64 mg at the time of harvest compared to 45 mg and
24 mg in the hydrochar and the corn silage treatment, respectively
(Table 2). Leaf area was also remarkably higher with 23 cm2 when
plants were amended with pyrochar (hydrochar: 9 cm2, corn silage:
6 cm2). Additionally, specic leaf area was signicantly higher
compared to the corn silage and the hydrochar treatment (Table 2).
The effects of the different soil types and fertilizer levels on these
latter parameters were not signicant. A noteworthy effect caused
by the presence of hydrochar was a larger heterogeneity of the
measured plant parameters compared to the two other treatments.
The root system of winter wheat showed remarkable and
consistent differences between the different treatments (Fig. 1).
Plants on soil amended with hydrochar had a very short root system, which was not reaching the bottom of the pots. Side roots
were short and had a stunted look. In contrast, plants on soils with
corn silage developed very long and ne roots concentrating at the
bottom of the pots. Plants on soils with pyrochar showed an
Table 2
Inuence of amendment types on various plant characteristics. Numbers of days are
given as days after sowing. The parameter Fv/Fm is an indicator of the quantum yield
of photosystem II. Different letters indicate signicant differences at a 0.05.
Plant characteristic
Type of amendment
Corn silage
75
91
6.6
15.3
16.2
18.9
24.4
1.1
0.69
5.7
292
25 b
11 b
2.3 b
4.5 a
4.4 a
5.2 a
7.4 a
0.3 a
0.05 a
1.8 a
28 a
Hydrochar
48
74
4.0
15.0
17.9
24.1
44.5
0.3
0.78
9.2
299
31 a
27 a
2.3 a
4.9 a
4.6 a
4.9 b
27.7 b
0.1 b
0.01 c
3.3 b
32 a
Pyrochar
77
93
7.5
17.8
23.6
18.9
63.6
0.4
0.73
22.5
400
22 b
12 b
3.0 c
4.3 b
5.9 b
6.1 c
5.6 c
0.2 b
0.05 b
5.1 c
43 b
269
270
Fig. 1. Typical root system of winter wheat plants grown with corn silage (left panel), hydrochar (middle) and pyrochar (right panel).
Fig. 2. Shoot N measured at the day of plant harvest. Results for the sandy soil are shown in the left half of the graph (shaded in grey) and results for the calcareous soil in the right
half. Numbers 0, 1 and 2 indicate the different fertilizer levels: no KNO
3 , 10 and 20 mg KNO3 per pot. A univariate analysis of variance revealed the signicant inuence of soil type,
char type and fertilizer level. Different letters and asterisks denote signicant differences at a 0.05. Data are given with respective standard deviations. HTC: hydrochar, PC:
pyrochar.
Table 3
Inuence of soil type, type of amendment, fertilizer level and plant presence on
selected plant and soil parameters according to univariate ANOVA. A factor is
considered to be of signicant inuence if p < 0.05.
Plant/soil parameter p-values for the inuence of
15
Fig. 3. Percentage of added KNO
N incor3 fertilizer used by plants as revealed by
poration. F1 and F2 denote the fertilizer levels (10 or 20 mg N pot1). Different letters
indicate signicant differences at a 0.05. Soil type had no signicant inuence,
squares give mean values of both fertilizer levels. Data are given with respective
standard deviations. HTC: hydrochar, PC: pyrochar.
Shoot N content
Fertilizer use
NO
3 at harvest
NH
4 at harvest
respired CO2
Microbial biomass C
Plant presence
0.000
0.073
0.027
0.910
0.293
0.003
e
e
0.000
0.002
e
0.001
0.000
0.000
0.000
0.000
0.000
0.000
0.001
0.200
0.026
0.987
0.103
0.297
271
1
large amounts of NH
dry soil) in the
4 (up to 0.15 mg NH4 eN g
soils amended with hydrochar came from and what happened with
the added fertilizer NO
3 . Both microbial biomass and microbial
respiration were relatively high in the presence of hydrochar. This is
Fig. 4. NO
3 and NH4 concentrations in soils after harvest of plants. NO3 concentrations were signicantly inuenced by the different soil types, char amendments and in the case of
pyrochar also by fertilizer level as well as the presence of plants. For hydrochar, values on calcareous soil are shown separately for pots with and without plants because there were
visible differences, however, these differences were not signicant due to a high variance. The square in between shows the mean value for pots with and without plants. NH
4
concentrations were only inuenced by type of amendment and the presence of plants (Silage, hydrochar). Data are given with respective standard deviations. HTC: hydrochar, PC:
pyrochar. Different letters and asterisks denote signicant differences between treatments at a 0.05.
272
Fig. 5. Soil respiration measured as CO2 evolution in 3 min (a) and microbial biomass carbon (b) in the different treatments. Fertilizer level had no signicant inuence on CO2
evolution, values were only determined for pots without plants, light brown parts of the bars show fraction arising from the decomposition of native soil organic matter. Microbial
biomass carbon was also independent of fertilizer level but signicantly inuenced by soil type, type of amendment and presence of plants. Here, green bars show results in the
presence of plants, dotted bars symbolize calcareous soil, plain bars sandy soil. Data are given with respective standard deviations. HTC: hydrochar, PC: pyrochar. (For interpretation
of the references to colour in this gure legend, the reader is referred to the web version of this article.)
273
Fig. 6. Summary of interactions between the three soil amendments and the nitrogen balance. The dotted line in the right panel symbols interactions between nitrate and the
surface of the pyrochar. Silage structure from: Yinghuai et al. (2013), hydrochar structure from: Kumar et al. (2011), pyrochar structure from: Heidenreich et al. (1968). See text for
further explanation.
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