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Soil & Tillage Research 163 (2016) 224234

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Soil & Tillage Research


journal homepage: www.elsevier.com/locate/still

The effects of birch (Betula spp.) biochar and pyrolysis temperature on


soil properties and plant growth
Marleena Hagnera,* , Riitta Kemppainenb , Lauri Jauhiainenb , Kari Tiilikkalab ,
Heikki Setla
a
b

University of Helsinki, Department of Environmental Sciences, Niemenkatu 73, 15340 Lahti, Finland
Natural Resources Institute Finland (Luke), 31600 Jokioinen, Finland

A R T I C L E I N F O

Article history:
Received 18 March 2016
Received in revised form 15 June 2016
Accepted 16 June 2016
Available online 1 July 2016
Keywords:
Birch wood biochar
Pyrolysis temperature
Ecotoxicity

A B S T R A C T

The addition of biochar to agricultural soils is recommended to improve soil functions and plant growth.
However, due to high variability in the quality of biochar, its effects on soils and plants are likely to differ.
We explored the impacts of pyrolysis temperature on the quality and usability of birch wood biochar as a
soil amendment. The impact of three biochar types pyrolysed at 300, 375 or 475  C on soil
characteristics and the growth of lettuce (Lactuca sativa), radish (Raphanus sativus), barley (Hordeum
vulgare) and ryegrass (Lolium perenne) was investigated in a greenhouse experiment. In addition, the
potential adverse effects of biochar on soil organisms (nematodes, earthworms, microbial biomass and
activity) were studied.
Biochar produced at the lowest temperature had initial transient negative effect on the germination
and biomass of lettuce, while biochar produced at higher temperatures had no such effect. After ve
weeks, pyrolysis temperature had only a marginal inuence on biochar-induced effects on soil pH, WHC,
soil organisms and plant growth. Our results suggest that birch biochar, irrespective of pyrolysis
temperature, has a parallel effect on plant biomass production and soil characteristics but the effect
depends on plant type and biochar application rate.
2016 Elsevier B.V. All rights reserved.

1. Introduction
Biochar, a carbon-rich material produced for example by the
slow pyrolysis of biomass, is suggested to be added to agricultural
soils to improve soil functions and to increase soil carbon
sequestration (Lehmann et al., 2003). During the past decade,
numerous articles focusing on the use of biochar have been
published, but have shown inconsistent results (Deenik et al., 2010;
Jeffery et al., 2011; Van Zwieten et al., 2010a). In general, the
addition of biochar to agricultural soils changes the physical,
chemical and biological properties of the soil. Biochar may enhance
plant growth, for example by increasing soil water-holding
capacity (WHC) and nutrient retention due to an increase in the
cation exchange capacity (CEC), or by improving physical
characteristics of the soil and mycorrhizal competence (Atkinson
et al., 2010). However, reactions in the soil after the addition of
biochar depend on the characteristics of biochar, soil, climate and
soil-inhabiting organisms. Biochar studies have mainly been

* Corresponding author.
E-mail address: marleena.hagner@helsinki. (M. Hagner).
http://dx.doi.org/10.1016/j.still.2016.06.006
0167-1987/ 2016 Elsevier B.V. All rights reserved.

performed in the tropics and temperate climates, but little is


known about the effects of biochar in colder climate, particularly in
the boreal zone (Tammeorg, 2014). In general, acidic soils and soils
with low organic matter content are assumed to benet from the
application of biochar via increasing soil pH and improved nutrient
retention and water holding capacity (Novak et al., 2009, 2012).
In addition to plant biomass production (Liu et al., 2013),
biochar is reported to affect other plant characteristics such as
nutrient content and resistance to diseases (Elad et al., 2010;
Nelissen et al., 2014b). Biochar addition has been reported to either
increase or decrease nutrient uptake by plants (Deenik et al., 2010;
Kloss et al., 2014; Lehmann et al., 2003; Nelissen et al., 2014b; Zhao
et al., 2014). Biochar in itself can provide nutrients for plant growth
or can increase nutrient bioavailability, for example by inducing
changes in soil pH and CEC (DeLuca et al., 2009). The reduced
nutrient availability for plants may be e.g. due to nutrient
adsorption to biochar or, in the case of nitrogen, due to enhanced
biotic N immobilization boosted by the labile C fraction in biochar
(Deenik et al., 2010; DeLuca et al., 2009). Some evidence suggests
that the water and soluble nutrients stored in biochar may be
available for plant uptake during, e.g. drought and nutrient

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

decient conditions (Cao et al., 2014; Taghizadeh-Toosi et al., 2012)


but there is a need for further evidences.
Pyrolysis temperature has a large effect on biochar characteristics: the higher the pyrolysis temperature, the higher the pH,
surface area, carbon content and stability of biochar (Angin and
Sensz, 2014; Keiluweit et al., 2010). At low temperatures, biochar
retains several chemicals and nutrients that are lost at higher
temperatures (Chan and Xu, 2009), has a less condensed Cstructure, higher volatile matter (VM) content (Chan and Xu, 2009;
Keiluweit et al., 2010) and thus greater reactivity in the soil.
Recently, several studies concerning the effects of pyrolysis
temperature on the physiochemical properties and structure of
biochar have been published (Angin and Sensz, 2014; Sun et al.,
2014). Temperature-induced changes on biochar properties may
have effects on soil properties (Nelissen et al., 2014a, b; Novak
et al., 2009) and plant response to biochar addition (Liu et al.,
2013). However, large scale investigations aiming at connecting
biochar pyrolysis temperature to the growth of several plant
species, soil parameters and -organisms are scarce.
The incomplete carbonization of biomass at low temperatures
can result in the formation of toxic organic substances, such as
polycyclic aromatic hydrocarbons (PAHs) and phenols, on biochar
surfaces (Hilber et al., 2012). For example, PAHs in biochar have
been reported to inhibit plant germination (Rogovska et al., 2011).
Besides plants, biochar can also affect soil biota for example
earthworms (Liesch et al., 2010; Tammeorg et al., 2014). Since
pyrolysis temperatures have a large impact on the formation of
toxic substances, it may be possible to reduce the toxic effects of
biochar in soils by selecting the right feedstock material and
suitable pyrolysis temperatures (Yargicoglu et al., 2014). The
pyrolysis temperature of traditional charcoal/biochar retorts varies
considerably between retorts and even within a retort. In Finnish
commercial slow pyrolysis retorts, the target temperature is
typically ca. 450  C (Fagerns et al., 2012), which is within the
limits given by the European Biochar Certicate (EBC, 2012) i.e.
350  C to 1000  C.
The main aim of this study is to increase our knowledge on the
impacts of biochar as a soil amendment, especially to explore the
impact of pyrolysis temperature on the quality and usability of
birch (Betula spp.) hardwood biochar as a soil amendment. Since
birch is a relatively fast growing, common and thus widely
available material for biochar production, birch wood provides a
possible feedstock material in northern latitudes. The rst
objective was to measure the impacts of three biochar types
pyrolysed at different temperatures on the growth of three
different crop plants. The second aim was to measure phytotoxicity
and the toxicity of biochar types to a selected group of soil
organisms. As earthworms are known to substantially affect soil
quality and plant growth (Edwards and Bohlen, 1996), the potential
impacts of biochar on these macrobiota is of fundamental interest.
We hypothesized that pyrolysis temperature affects the responses
of biochar in soil in the following manner:
1. Biochar increases soil water holding capacity and pH, more so at
higher pyrolysis temperatures
2. Pyrolysis temperature has an effect on the quality of biochar in
terms of affecting nitrogen availability for plant uptake
3. Biochar produced at low temperatures retains toxic substances
and has a negative effect on (i) plant performance and (ii) the
survival and behavior of earthworms
4. Biochar produced at higher temperature increases plant growth
more than biochar produced at lower temperature
This study forms part of a larger research program in which
variation in the pyrolysis process temperature on the composition

225

and chemical characteristics of biochar were investigated (Fagerns et al., 2014; Hagner et al., 2015).
2. Materials and methods
2.1. Biochar, soil and plants used
Three types of biochar derived from birch wood (Betula spp.)
were produced by the VTT Technical Research Centre of Finland,
using a pilot-scale slow-pyrolysis unit. Birch branches with
moisture content of 1012% were cut to 25 mm  50150 mm
pieces. A temperature prole with two phases and overall duration
of 7.5 h was used to produce biochar. Temperature was rst raised
to 280  C (3.5 h) and thereafter to nal temperatures of 300  C,
375  C or 475  C (holding time 4 h). The biochar types are identied
according to the nal temperature as BC300, BC375 and BC475.
Pyrolysis temperatures 375  C and 475  C were within the range of
the European Biochar Certicate (EBC, 2012) and typical in Finnish
pyrolysis retorts. The lowest pyrolysis temperature (300  C) was
chosen to study if temperature lower than the one suggested by the
EBC would result in lower biochar quality and usability in the eld.
Volatile matter (%) (SFS-EN 15148), ash content (SFS-EN 14775),
xed carbon (ASTM D 3172), C/H/N- (SFS-EN 15104) and S-content
(SFS-EN 15289) were analysed according standard methods. PAHconcentrations were analysed in Nab Labs Ltd (GC/MS (SIM), ISO
16703). The BET surface area (Standard PANK 2401), water holding
capacities (WHC), pH (v/v 1:5 biochar:distilled water) and other
characteristics of the biochar types are presented in Table 1. The
pyrolysis process and the characteristics of the biochar types were
analysed at VTT Technical Research Centre of Finland and are
reported in detail in Fagerns et al. (2014).
A sandy-silt soil (classied as Cambisol based on Finnish and
FAO classication systems) was shoveled (ca. 10 cm deep surface
layer) from an organic agricultural eld in Rehtijrvi, SW-Finland
in the autumn of 2012. The moisture content of the soil, collected
after a very dry period, was 1.0% (Standard ASTM D2216, ASTM
2010). The soil consisted of 22% medium sand (200600 mm), 57%
ne sand (60200 mm) and 21% of particles under 60 mm (silt and
clay) in size. Soil nutrient analyses (Ca, K, Mg, P) were conducted in
the laboratory of the MTT Agricultural Research Finland according
to the protocol by Vuorinen and Mkitie (1955) used widely in
routine soil testing in Finland. The soil had a pH of 6.02 (1:5 soil:
water v/v), electrical conductivity of 0.68 mS/cm and pore volume
of 47% (Sandbox-method, Eijkelkamp Agrisearch equipment
2013). In general, soil was quite poor as concentrations of Ca, K, Mg,

Table 1
Characteristics of biochar types produced at pyrolysis temperatures of 300  C
(BC300), 375  C (BC375) and 475  C (BC475). Adapted from Fagerns et al. (2014).
Biochar property

Ash content (%)


Volatile matter (%)
Fixed carbon (%)
C (%)
H (%)
N (%)
O (%)
S (%)
Surface area (m2/g)
WHC (%)a
pH (1:5 H2O)
PAH (mg/kg)
Bulk density (g/cm3)b

Biochar type
BC300

BC375

BC475

0.5
48
52
72
4.9
0.2
23
0.01
2.2
23
5.1
194
0.41

0.7
30
69
80
3.9
0.3
15
0.01
6.4
24
5.2
4100
0.41

1.0
17
82
89
3.1
0.3
7
0.01
44
288
7.5
1107
0.45

a
WHC: water holding capacity (24 h) i.e. the total amount of water a material can
hold (% of biochar mass).
b
Bulk density (g/cm3) = dry mass (g)/volume (cm3).

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M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

and N were 760 mg/kg, 93 mg/kg, 34 mg/kg and 0.45 mg/kg,


respectively. However, P concentration was high (23 mg/kg).
Earthworms, larger stones and roots (length >10 mm) were
removed from the soil prior to lling it, into the experimental
pots (see below). Before using in the experiment (see below) the
moisture content of the soil was adjusted to 26% (ca. 50% of WHC).
Lettuce (Lactuca sativa) was used as test plant to study the
inuence of biochars on germination and initial growth of the
seedlings. Radish (Raphanus sativus), barley (Hordeum vulgare) and
ryegrass (Lolium perenne) were used as test plants to model the
cycle of crop rotation of three plant families. Lettuce seeds (cultivar
Australishe gele NO:8281) were obtained from Econova Garde ab,
radish seeds (cultivar Poker) from Helle Oy (21410 Vanhalahti,
Finland) and seeds of barley (cultivar Voitto) and ryegrass (cultivar
Riikka) from Boreal Plant Breeding Ltd. (31600 Jokioinen, Finland).
2.2. Greenhouse experiment
2.2.1. Establishment of the pot experiment
A study was performed in a greenhouse at the MTT Agrifood
Research Finland in Jokioinen, in 2013. The experiment was
conducted in 1.5 L owerpots ( 11 cm, height 19 cm) with four
holes ( 0.5 cm) at the bottom. The biochar produced was passed
through a 2 mm sieve before addition to the soil. The treatments,
each with seven replicates, consisted of soil (1 L = 1320 g, moisture
content 26%) mixed with 1) 20 g of either BC300, BC375 or BC475,
2) 80 g of either BC300, BC375 or BC475 or 3) a control system
without biochar. The application rate of biochar to the pots
corresponded to an application rate of 20 t/ha or 80 t/ha in the top
10 cm depth. The experimental design consisted of two identical
and simultaneously-conducted pot experiments on four tables,
each with 28 pots, altogether 112 pots (Fig. 1). On each table the
four treatments were assigned to 28 pots according to the rowcolumn experimental design. This design is useful in greenhouse
experiments because it takes into account two-dimensional
gradients, while a randomized complete block design only
considers one dimension (Williams et al., 2002). The biochar
was manually mixed (5 min) with the soil, separately for each pot.
After lling the pots, the soils (all pots) were fertilized with 25 mL
of a 1.8% dilution of Yara Combi 1 (NPK 14-11-25). An exclusion area

extra pots growing ryegrass was established around the


experimental pots to prevent edge effect (Fig. 1). The experiment
ran for 30 weeks (9/2012-3/2013) and constituted of ve phases: 1)
stabilization period (5 weeks), 2) radish growing season (5 weeks),
3) barley growing season (6 weeks), 4) winter rest (4 weeks) and 5)
ryegrass growing season (10 weeks). As numerous factors can
affect grain yield production of cereals in a greenhouse pot tests,
the aim of the separate growth periods for the plants was to
explore only the effects of biochar types on plant biomass
production, not grain yield.
2.2.2. Plant growth trials
During the stabilization period the pots were kept in the
greenhouse with a light/dark cycle of 16/8 h and day/night
temperatures of 20/15  C. The pots were randomly placed on
plates on a moist lter bed to ensure constant air moisture during
incubation. During the stabilization period, the germination
inhibition assay with Lactuca sativa was examined. Three days
after lling the pots, 10 lettuce seeds were sown in each pot (n = 14/
treatment). The soil surface was covered with a plastic lm for
2 days to maintain stable soil moisture conditions. After seed
germination, the plastic lms were removed. Additional irrigation
was performed by spraying the soil surface daily. Germination of
the lettuce seeds was checked 14 days after sowing. The seedlings
were uprooted and dried for the analysis of dry mass and nitrogen
content (LECO CNS-analyser). After the germination inhibition
assay the moisture content of the potted soils were monitored
weekly using the Grodan WCM control meter. If soil water content
(WC) decreased below 20%, extra irrigation (ca. 50 mL three times a
week) was added manually to each pot to keep WC -values of soils
at ca. 2025% (ca.% of WHC).
After the stabilization period, 10 radish seeds were sown in the
same pots from which the lettuce seedlings were removed
(n = 14/treatment). The plates under the pots were removed.
Moisture in the pots and humidity around the pots were stabilized
with a lter tissue kept wet under the pots during the duration of
the experiment. The temperature in the greenhouse was day/night
20/15  C and the light/dark cycle was 16/8 h. The pots were covered
with plastic lms for 5 days to maintain constant soil moisture
conditions during germination. Irrigation was applied with an

Fig. 1. The greenhouse experimental design. Letters A-D represent the four tables on which the experimental pots resided. Numbers represents the pots on the tables:
1 = Control, 2 = BC300, 3 = BC375 and 4 = BC475, and in an exclusion area (5) around the primary exam pots. For tables A and B, the biochar application level was 20 g/L and for C
and D, 80 g/L of soil.

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

automated drip system; 35 mL twice a day. After 11 days, the crop


was thinned to the three best seedlings per pot. The condition of
the seedlings was estimated visually (discoloration/no discoloration) and dry mass and nitrogen content of the harvested
seedlings were analysed. The remaining radish seedlings were left
to grow for 25 days before the analysis of their wet and dry mass.
Leaf and root biomass of the plants were weighed separately. After
removing the radish plants and to simulate normal tilling
practices the soil was removed from the pot, mixed properly,
fertilized (50 mL, 1.8% Yara Combi 1) and put back into the same pot
on moist lter bed. On the next day, 10 barley seeds were sown in
each pot. The pots were covered with a plastic lm for 4 days to
promote germination. The temperature in the greenhouse was
adjusted to 16/10  C (day/night) and a light/dark cycle of 16/8 h,
with irrigation of 35 mL twice a day. The seedlings were staked
once they reached a height of 15 cm. Barley plants were left to grow
for 6 weeks before the analysis of dry mass.
An articial winter (rest period) was introduced after the
harvesting of barley. The pots were covered with brown craft paper
and kept at 10  C without light at constant air humidity (45% RH)
for four weeks. At the start of the last phase, the soils were again
removed from the pots, mixed properly, fertilized (50 mL, 1.8% Yara
Combi 1) and put into 1 L pots as the amount of soil decreased
because of soil sampling (see below). Temperature of the
greenhouse was adjusted to 18  C. Seeds of Lolium perenne were
sown in the pots (ca. 150 seeds, 0.305 g/pot) and then covered with
a plastic lm for 4 days. The pots were placed on moist lter bed. In
addition, the soils were sprayed daily (7 days) with water until the
seeds germinated. After germination, the irrigation was applied
with an automated drip system twice a day (1023 mL). During a
10-week period, the yield was harvested six times (3, 4, 5, 7, 8 and
10 week after ryegrass was sown) to measure plant dry mass.
During the 10 weeks of growth, pots were not fertilized in order to
test whether nutrients absorbed to the biochar were available to
the plants. Temperature and relative humidity on the experimental
tables were measured with two Wiseman Klein WK057 data
loggers. Effective temperature sums (ETS) were calculated for each
crop from the sowing day to the harvesting day. The threshold
value was +5  C. ETS values in day degrees were 457 for radish, 471
for barley and 295, 160, 112, 196, 147 and 144 for the six harvesting
times of the ryegrass.
2.2.3. Soil pH, basal respiration and nematodes
Several soil parameters were assessed during the study. To
minimize the amount of soil loss from the pots during the
experiment, soil samples were taken only three times: at the end of
the stabilization period (Week 5, since the start of the study), the
radish growing season (Week 10) and at the end of the experiment
(Week 30). During each soil sampling event three soil samples
were taken from each pot using a corer ( 1.5 cm, 5 cm deep, ca.
9 mL). Soil samples were stored (15 days) at 5  C for the analyses
(the exception: 80 g/L samples for the analysis of microbial activity
were stored at freezer at 18  C, max two months). Water content
of the soil samples was determined after drying ca. 5 g of soil
samples at 105  C for 24 h. Soil pH was measured from samples
taken at the end of the radish growing season (Week 10) in a 1:2.5
(v/v) soil:distilled water suspension according to the ISO 10390
standard (ISO, 2005).
Microbial activity was measured from soil samples taken from
pots residing on tables A (20 g/L) and C (80 g/L) whereas nematodes
were extracted from pots on tables B (20 g/L) and D (80 g/L) (n = 7/
treatment). This restriction was necessary to minimize soil loss.
Soil microbial activity was measured from 20 g fresh soil using an
Apollo 9000 Total Organic Carbon (TOC) analyzer modied to
analyze head-space air samples in the Almalab (University of
Helsinki). Roots were removed from the soil before measurement.

227

The soil was placed in a 43 mL glass jar, and then allowed to


stabilize for 24 h (21  C) before the rst measurement: a 0.5 mL air
sample was taken from the head-space of the jar through a rubber
lid using a syringe. A second air sample was taken two hours later.
Basal respiration was calculated based on the difference in the
amount of CO2 between the two measurements. Abiotic CO2
production of biochar was measured by placing 10 g of each
biochar type and 5 mL water in a glass jar and let the mixture
stabilize for 24 h at room temperature (n = 3). The rst and second
measurements and calculations were done as above.
Nematodes were extracted from 15 g of fresh soil samples (n = 7)
using the wet funnel method (Sohlenius, 1979). The nematodes
extracted were counted and functional groups determined under a
binocular microscope (50 individuals/sample) according to their
food-specicity (plant-, fungal-, bacterial feeding, omnivorous,
animal predators) (Yates et al., 1993).
2.3. Earthworm studies
2.3.1. Acute toxicity test
Earthworms (Eisenia fetida) used in the test were obtained from
cultures maintained at the MTT Agrifood Research Finland. Prior to
the toxicity test, the worms were maintained in plastic boxes
(50  80  30 cm) with a 5 cm layer of the mixture (v/v 1:1) of
sphagnum peat and commercial garden soil. pH of the mixture was
adjusted to 6.0  0.5 with CaCO3. The worms were fed weekly by
adding a 5 cm layer of horse manure in the containers. The manure
was kept frozen for two weeks before use to eliminate insect eggs
and larvae. Earthworms were transferred into the articial test soil
one day before the tests started.
The toxicity of biochar (BC300, BC375, BC475) to E. fetida was
determined according to the OECD Guidelines Earthworm, Acute
Toxicity Tests 207 (OECD 1984). Articial soil containing 10%
Sphagnum peat (sieved through 4 mm), 20% kaolin clay and 70%
quartz sand was prepared. CaCO3 was used to adjust pH of the soil
to 6.0  0.5. Biochar was crushed manually and sieved to <2 mm.
Biochar was then mixed with dry soil constituents (manually for
5 min) to achieve biochar concentrations of 0, 21, 35, 60 and 102 g/L
(soil dry mass). Moisture contents of the nal test soils were
adjusted to 70% of WHC using Milli-Q water by following the
protocol by Li et al. (2011). The amount of added water uctuated
between 116 (control) and 158 mL (BC475 at 102 g/L). After adding
750 g of moist soil to transparent plastic containers (130 mm
 100 mm  70 mm), ten mature (judged by the presence of
clitella) earthworms were placed on the soil surface of each
container. Once the worms entered the soil, the container was
covered with a lid. This procedure was conducted simultaneously
for each of the three types of biochar, each with 4 replicates. The
containers were placed in a controlled environment chamber at
20  C under constant illumination. After 7 and 14 days living
worms were counted.
2.3.2. Earthworm avoidance study
The avoidance behavior of earthworms (E. fetida) to biochar was
studied according to Van Zwieten et al. (2010b). Articial soil was
prepared as described above. Biochar was crushed manually and
sieved to below 2 mm. The used biochar concentration was 12 g/L
dry weight corresponding to 12 t/ha (relevant application rate in
the eld use). Instructions by Li et al. (2011) was followed to
achieve adequate wetting of the soil and biochar-soil blend for the
earthworm survival. Water contents were adjusted to 70% WHC
separately for each treatment. The test procedure was conducted
simultaneously for each biochar type with ten replicates.
Transparent plastic containers (170  100  80 mm) were divided into two halves with a plastic split. Amended soil (400 g) was
placed on one side and nonamended soil on the other side of the

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M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

container. To test that worms were homogenously distributed in


the containers an extra set of containers was constructed with
control soil only. The divider was then removed and ten mature
worms were placed in the middle of the containers. Once the
worms entered the soil, the container was covered with a
perforated lid. The containers were placed in a controlled
environment chamber under constant illumination at 20  C. After
2 and 7 d, the split was reintroduced in the marked position and the
number of individuals in the amended and unamended halves
counted.

there were no differences in soil pH between the treatments. At the


highest application level (80 g/L), the decrease of soil pH was lower
in biochar treated soils than in control soil with nal pH being 5.04
in BC300, 5.09 in BC375 and 5.35 in the BC475 treated soil. During
the rst weeks of the experiment, moisture content of control soils
remained higher than in biochar treated soils (p < 0.05). After the
winter rest (weeks 1620), no statistically signicant differences in
soil moisture values existed between biochar treated and control
soils (p > 0.05), except at Week 20 when the moisture content of
biochar treated soil was higher than in control soil (Fig. 2).

3. Statistical analysis

4.1.2. Germination and nitrogen uptake by test plants


At the lower application level, none of the biochar types
affected germination, dry weight or nitrogen content of L. sativa
seedlings (p > 0.05 in all cases). However, at the higher application
level, BC300 inhibited L. sativa germination (Table 2) (p = 0.001),
and BC300 and BC375 reduced the dry-weight of L. sativa seedlings
statistically signicantly (p = 0.001, p = 0.003, respectively). Biochar
types (20 or 80 g/L) had no effects on N concentrations of L. sativa
seedlings; except the BC300 at 80 g/L which reduced N concentrations signicantly (p = 0.004) compared to control (Table 2).
None of the biochar types had an impact on the germination of
radish seeds (p > 0.05 in all cases). Instead, each biochar type
increased the biomass of radish seedlings 11 days after sowing
(p < 0.05 in all cases) (Table 2) with no differences between
biochars. At the lower application level, each biochar type induced
ca. a 43% increase and at 80 g/L ca. 84% increase in the biomass of
radish seedlings (Table 2). Nitrogen concentrations of the seedlings
decreased signicantly when the plants were growing in BC375
and BC475 (p = 0.044, p = 0.006, respectively) amended soils of both
application rates (Table 2). Seedlings in the soils treated with
BC300 and BC375 showed ca. 68% and 64% less discoloration than
the control seedlings, respectively (rates analysed together:
p = 0.004, p = 0.009, respectively) (Fig. 3). Radish seedlings growing
in the BC475 treated soils showed an average 44% decrease in
discoloration when compared to the control but this result was not
statistically signicant (p = 0.065) (Fig. 3).

The statistical model used for the analysis of plant growth,


germination and N-content was based on the row-column
experimental design. The model was:
yijklm m leveli tablelevelij rowtableik columntableil
treatmentm treatment  levelim eijklm
where m is the intercept, leveli, treatmentm and treatment*levelim
are xed effects of the biochar application level (i = 20 or 80 g/L),
treatment (m = Control, BC300, BC375, BC475) and their interaction, respectively. Table(level)ij, row(table)ik, column(table)il and
eijklm are the effects of the table (j = two tables for both biochar
application levels), row (k = 1, 2, 3, 4), column (l = 1, . . . ,7) and the
residual error. If differences between treatments were similar for
both biochar application levels, results were showed as the main
effect of treatments (14 replicates/treatment) and Tukey-test was
used for multiple comparisons of treatments. The underlying
distribution of the radish discoloration data was binomial. Data
were analysed using a generalized mixed model with the logit-link
function and binomial distribution, using the SAS/GLIMMIX
software.
To examine differences in soil micro-organisms and nematodes
between treatments, a repeated measurement ANOVA was
conducted with time and biochar as factors (SPSS, 2012). As
expected, there was an interaction between time and biochar, i.e.
the effects of biochar treatments differed between sampling
events, most likely due to the aging of biochar (e.g. oxidation and
degradation processes). Even though the different measurement
times were not independent statistical units, the effects of biochar
at sampling events were analysed separately using one-way
ANOVA. Therefore the statistical signicances between biochar,
nematodes and micro-organisms should be considered with
caution.
Avoidance percentage in the earthworm avoidance test was
calculated according to Amorim et al. (2005):


nc  nt
 100
Xavoid
n

4.1.3. Plant growth responses


Irrespective of treatment, radish, barley and ryegrass grew well
during the experiments. After the growth periods, radish plants
had reached maturity with well-developed bulbs, ryegrass shoots
reached the height of ca.15 cm six times (harvesting was done
when the shoots achieved 15 cm height) and barley shoots were ca.

where Xavoid is the avoidance percentage, nc is the number of


worms in the control soil (mean of ten replicates), nt is the number
of worms in the bioichar-treated soil (mean of then replicates), and
n is the total number of earthworms at the beginning of the
experiment. Differences between control and biochar treatments
were analysed using a Paired Samples T-test at 95% signicance
level (SPSS, 2012).
4. Results
4.1. Greenhouse experiment
4.1.1. Biochar effects on soil pH and moisture
The pH of the control soils decreased during the experiment
from 6.02 to 4.73 (Week 10). At the lower application level (20 g/L)

Fig. 2. Mean ( SE) water content (measured using the Grodan WCM control meter)
of the soils in the biochar (80 g/L) treated pots (BC350, BC 375 and BC475 pooled)
and in the controls without biochar amendment (n = 14/treatment). At the
beginning of the experiment (Week 0), before the sowing of radish (Week 5),
before the sowing of barley (Week 10), before the sowing of ryegrass (Week 20) and
after ryegrass was cut (Weeks 25, 26, 28, 29, 30). Statistically signicant differences
to the control are marked with an asterisk * (p < 0.05).

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

229

Table 2
Germination, biomass and nitrogen content of the lettuce and radish seedlings 14 and 11 days, respectively, after being sown (mean, n = 14). Statistically signicant differences
compared to the control are marked with an asterisk (* p < 0.05; ** p < 0.01).
20 g/L

80 g/L

Plant species

Treatment

Germinated seedlings

Biomass (mg dw/pot)

N (%)

Germinated seedlings

Biomass (mg dw/pot)

N (%)

Lactuca sativa

Control
BC300
BC375
BC475
Control
BC300
BC375
BC475

9.2
9.2
9.4
9.6
9.5
9.6
9.6
9.7

37.3
35.2
35.1
36.9
147.8
212.7**
215.4**
219.9**

6.4
6.4
6.6
6.8
7.7
7.6
7.4*
7.3**

9.1
5.9**
8.7
8.7
9.1
9.8
9.1
9.6

34.5
16.4**
27.2**
30.2
109.1
200.5**
208.4**
226.1**

6.4
5.7**
6.0
5.9
8.0
7.7
7.6*
7.2**

Raphanus sativus

Fig. 3. Effects of the three biochar types (BC300BC475) on the amount (%


individuals) of radish seedlings displaying discoloration on the leaves 11 days after
germination (mean,  SE, n = 14). Statistically signicant differences compared to
the control (without biochar addition) are marked with an asterisk (* p < 0.05; **
p < 0.01).

20 cm high. The lower biochar rate of 20 g/L had no effect, on the


biomass of radish leaves (p = 0.730) while at the higher application
level (80 g/L), BC375 and BC475 increased the aboveground
biomass of radish signicantly (p = 0.001, p = 0.002, respectively).
BC300 (80 g/L) had no effect on the biomass of radish leaves
(p = 0.094). All biochar types signicantly stimulated radish root
growth at both application levels (p < 0.001 in each case) with no
differences between the biochar types (Fig. 4).
None of the biochar types and application rates inuenced the
yield of barley (p = 0.212). The effects of biochar on the biomass of
ryegrass varied during the study. The lower application rate had no
signicant effects on ryegrass growth, except for the last harvest

Fig. 4. Root biomass (g dry weight/pot) of radish after 5 weeks of growth in the
control and biochar (BC300, BC375 and BC475) treated pots (mean  SE, n = 14).
Statistically signicant differences to the control are marked with an asterisk
** (p < 0.01).

when yield in the control pots was signicantly lower than in the
BC300 and BC475 treated pots (p < 0.05). At the higher rate, the
effects became evident: for the rst harvest (3 weeks after sowing)
each biochar type reduced ryegrass biomass signicantly (0.34 g in
control pots and 0.19, 0.25 and 0.30 g in the BC300, BC375 and
BC475 treated pots, respectively, p < 0.005 in all cases) (Fig. 5).
Biomass in the BC300 treated pots was signicantly lower than in
the BC375 or BC475 treated pots (p < 0.05). During the study the
effect of biochar changed and at the last harvest, the yield was
0.87 g in control pots and 1.22, 1.35 and 1.29 g in the BC300, BC375
and BC475 treated pots, respectively (p < 0.001) (Fig. 5). When
ryegrass dry mass during the study period was analysed
cumulatively, all soils treated with biochar (BC300, BC375,
BC475) produced greater yields than the control (p < 0.001). There
were also differences between the biochar types: BC300 produced
lower yields than BC375 and BC475 (p < 0.01).
4.1.4. Effects on soil organisms
At the low application rate, none of the biochar types affected
soil microbial activity. However, at the higher rate, BC300 and
BC375 increased soil microbial activity ca. 42 and 46%, respectively
(p < 0.001 in both cases) ve weeks after placing in the soil
(Table 3). Ten and thirty weeks after mixing the biochar in the soil,
microbial activity did not differ between treatments (p > 0.05 in all
cases). In the separate laboratory study, each biochar types
released CO2 after distilled water was added to the dry biochar.
The amount of CO2 released varied signicantly between biochar
types, being 0.84 in BC300, 1.06 in BC375 and 0.31 mg C/g/h in
BC475 (p < 0.05 in each case). When comparing the transient
abiotic CO2 release from the pure biochar to the CO2 release from

Fig. 5. Cumulative biomass of ryegrass (g dry weight/pot) in the control, BC300,


BC375 and BC475 treated pots at separate harvests (3, 4, 5, 7, 8 and 10 week after
sown) at 80 g/L biochar addition (n = 14). Cumulative biomass of the columns with
different letters differ signicantly (p < 0.05).

230

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

Table 3
Mean (SE) microbial activity (mg C g1 dm soil h1) in biochar (BC300, BC375 and
BC485) treated pots at application rates 20 and 80 g L1 and control soils without
biochar amendment. Statistically signicant differences to the control are marked
with a (p < 0.01). Microbial activity of soils that received 20 g L1 biochar were
analysed within a week after sampling. Soils with 80 L1 biochar were stored in a
freezer (max. 2 months) until being analysed.

20 g L1

80 g L1

Control
BC300
BC375
BC475
Control
BC300
BC375
BC475

Week 5

Week 10

Week 30

0.32  0.02
0.27  0.02
0.32  0.07
0.22  0.01
0.14  0.01
0.24  0.04
0.26  0.05
0.15  0.01

0.46  0.09
0.50  0.05
0.49  0.04
0.46  0.04
0.39  0.05
0.44  0.03
0.50  0.05
0.32  0.03

1.32  0.07
1.33  0.06
1.48  0.07
1.56  0.12
0.76  0.03
0.64  0.04
0.58  0.07
0.55  0.10

a
a

the soil samples taken at weeks 5, 10 and 30, the abiotic release
corresponds to ca. 1.5-5% from the observed total CO2 release.
However, as the actual abiotic CO2 release from the biochar types at
5, 10 and 30 weeks after biochar application in the soil is not
known, the attribution of the observed degradation of biochar/CO2
release after biochar addition to abiotic causes is difcult. Thus the
abiotic release of CO2 was not taken into account in the microbial
activity calculations.
At the rst sampling event (Week 5) the number of nematodes
in soils varied between 3.3 and 5.6 individuals/g soil (dm). The
population of nematodes increased during the study being 9.717
and 3548 individuals/g (dm) at Week 10 and Week 30
(respectively) with no differences between control and biochar
treated soils (p > 0.05) (Table 4) or different biochar rates (20 g/L
and 80 g/L). Similarly, none of the biochar types inuenced the
functional group composition of the nematode community. At
Week 5, herbivores were the most prominent functional group. At
Week 10, bacterivores became more frequent, followed by
herbivores and fungivores (Table 4).
4.2. Earthworm studies
4.2.1. Earthworm toxicity test and soil WHC
Biochar types had no toxic effects on the earthworm Eisenia
foetida as deduced from mortality rates of the worms even in the
highest used concentration 102 g/L. As the mortality of earthworms during the 14 days test was <2% in control treatment, the
test met the validation criteria.
Biochar, irrespective of type, had a clear positive effect on soil
WHC. On average, 102 g/L of biochar application increased soil
WHC from 26% in the control soil to 36% in the biochar treated soils
(Fig. 6). The effect of biochar types on WHC of the treated soils did
not differ from each other.

Fig. 6. Effects of biochar application rate (x axis) and biochar types produced at
different temperatures (BC300BC475) on articial soil WHC in earthworm
avoidance study. Control columns (white bars) represent the containers without
biochar addition.

4.2.2. Earthworm avoidance test


BC300 and BC375 had no effect on the placing of earthworms in
the test containers. Instead, 2 days after mixing biochar in the soil,
BC475 tended to repel earthworms (avoidance percentage 28%,
p = 0.083) but after 7 days the effect was opposite, i.e. BC475
attracted (23%, p = 0.045) the worms (Fig. 7). As the distribution
of earthworms in the control containers was homogenous and no
earthworm mortality was observed during the study, the
earthworm avoidance test met the validation criteria.
5. Discussion
5.1. Effect of biochar on soil characteristics
The application of biochar to arable soils has been shown to
increase soil pH, WHC and nutrient retention capacity (Atkinson
et al., 2010). We hypothesized that birch wood biochar produced at
high pyrolysis temperatures should affect soil WHC and pH more
than biochar produced at low temperatures. This assumption was
based on earlier observations which showed that, biochar prepared
at higher pyrolysis temperature has a larger surface area and
higher pH than low-temperature biochar (Keiluweit et al., 2010;
Angin and Sensoz, 2014). Corroborating earlier ndings, the BETsurface area as well as the pH of biochar applied in our study clearly
increased with increasing pyrolysis temperatures (Table 1). The pH
of the control soils decreased (6.02 ! 4.73) during the experiment
probably due to the acidic fertilizer applied (Barak et al., 1997). At
low application rate (20 g/L) biochar did not affect soil pH. At the
higher application rate (80 g/L), pH of the biochar treated soils
remained higher than in the control soils, suggesting that biochar
may buffer soil against pH decrease (Ren-kou et al., 2012).

Table 4
Mean number (SE) of nematodes and the proportion (%) of various feeding groups in the biochar treated (BC300, BC375, BC475) and control pots at Weeks 4, 10 and 30 at
biochar application rate 80 g/L.
Individual g
Week 5

Week 10

Week 30

Control
BC300
BC375
BC475
Control
BC300
BC375
BC475
Control
BC300
BC375
BC475

3.3  0.6
3.3  0.6
5.6  1.2
4.4  0.8
9.7  2.7
17.6  5.5
14.4  2.6
15.5  3.0
35.9  4.6
39.8  14.3
37.8  12.0
48.0  13.7

1

dw

Bacterivore

Herbivore

Fungivore

Omnivore

Predator

28.3%
35.9%
41.2%
36.6%
85.3%
92.0%
92.5%
93.3%
86.2%
83.1%
83.5%
94.8%

63.4%
52.1%
51.1%
45.5%
12.3%
6.4%
5.4%
5.2%
7.7%
15.7%
13.9%
5.2%

8.3%
12.0%
7.1%
17.1%
2.1%
1.6%
1.8%
1.6%
6.2%
1.2%
2.5%
0.0%

0.0%
0.0%
0.5%
0.8%
0.3%
0.0%
0.3%
0.0%
0.0%
0.0%
0.0%
0.0%

0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%
0.0%

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

Fig. 7. Distribution of earthworms (E. foetida) in the differentially-treated halves of


the test containers after 2 and 7 days in the earthworm avoidance experiment. In
the containers, one side was lled with control soil and the other with biochar
(BC300, BC375 or BC475) treated soil. The columns represents means of the
treatment (n = 10). The black bars (biochar application) below the horizontal line
indicate earthworm avoidance, the ones above the line earthworm preference.
Statistically signicant differences to the control are marked with an asterisk *
(p < 0.05).

However, substantial application rates of birch-wood based


biochar are required to produce liming effects in sandy soils. As
hypothesized, differences existed between the biochar types in
that the BC475, having the highest pH, also buffered of the
experimental soil more efciently than biochar types produced at
lower temperatures. Differences in soil acidity between treatments
were smaller than 0.5 pH units in all cases. Interestingly, despite
the clear differences in WHC and BET surface areas of the biochar
types produced at different temperatures (Table 1), this did not
translate into differences in WHC between the biochar-amended
soils. These results suggests that, despite the high variation in the
characteristics of biochar, the effect of different biochar types on
soil characteristics (e.g. pH and WHC) may be quite similar. Thus,
the effects of biochar in agricultural soils, whether positive or
negative, are conditional not only on the biochar characteristics
(e.g. structure and functional groups, BET-area, pH and WHC) (Xie
et al., 2015), application rate and time of application (Ajayi et al.,
2016) but also on the physical-chemical characteristics of the soils
(e.g. soil nutritional status, pH, WHC) in which biochar is added
(Ajayi et al., 2016; Deb et al., 2016; Liu et al., 2013).
5.2. Toxicity studies
As hypothesized, low-temperature biochar had an initial
negative effect on the germination (BC 300) and biomass
(BC300, BC375) of plants, yet biochar produced at higher temperatures (BC475) did not have such effects. In line with our study,
Bargmann et al. (2014a) showed that plant-based biochar
pyrolysed at high temperatures (800860  C, 0.5 h) had no effect
on plant germination. Instead, Bargmann et al. (2014a, b) reported
severe germination inhibition of barley by low temperature
(190  C) produced hydrochar (a solid product of hydrothermal
carbonization of biomass), but the effect was of short duration and
the second round of cultivation was not affected. In line with
Bargmann et al. (2014b), phytotoxicity was apparently transient in
the soil, since biochar affected the rst cultivar (lettuce) but not the
subsequent cultivars (radish, barley and ryegrass). This suggests

231

that some phytotoxic compounds remained in the low temperature pyrolysed biochar in our study. Pyrolysis temperature may
affect, for example, the content of volatile organic compounds
(VOC) of biochar. Charcoal with high VOC content can show
negative effects on plant growth (Buss and Masek, 2014; Deenik
et al., 2010). Consequently, differences of VOCs in biochar can
contribute, at least partly, to the often observed differences in their
phytotoxicity (see Spokas et al., 2011). Our results support previous
ndings of e.g. Buss and Masek (2014) and Deenik et al. (2010) as
BC300 with the highest VOC content reduced lettuce germination
and/or growth more than birch wood biochar pyrolysed at higher
temperatures. We assume that some phytotoxic C compounds
remained in biochar produced at lower temperatures but that
these compounds are quickly volatilized in the air or degraded by
soil microbes. Since biochar containing VOC and other dissolved
organic compounds (DOC) that serve as a labile C source to soil
microbes, it is also possible that some of the N was incorporated
into the microbial biomass and was thus not available to plants,
which then decreased plant germination and growth (Deenik et al.,
2010). This conclusion is supported by increased microbial activity
(Week 5) in the soils applied with BC300 and BC375, containing
more labile C than BC475. Consequently, a stabilizing period of a
couple of weeks between the addition of biochar and the sowing of
seeds may help to reduce the phytotoxicity of biochar on plants.
It is also possible that lettuce is more sensitive to some
compounds or effects of biochar than the other plants used in the
current experiment. This warrants a new test with lettuce regrown
in the same soil. Furthermore, PAHs in biochar have been reported
to inhibit plant germination (Rogovska et al., 2011). However, in
our study, PAH concentrations of biochar did not correlate with the
germination of lettuce or radish. An obvious reason for this is that
all the tested biochar types had low total concentrations of PAHs
(Table 1). It is to be noted that the used biochar concentration
(80 g/L) showing phytotoxic effects of plants corresponds to 80 t/ha
at tillage depth of 10 cm, which is considered unrealistically high in
agricultural use.
Contrary to our hypothesis, none of the biochar types had an
effect on the survival of the earthworm E. fetida even at very high
concentrations (102 g/L). Liesch et al. (2010) also showed that
plant-based biochar had no effect on the survival of this
earthworm species. Interestingly, at the start of our experiment,
avoidance behavior increased with increasing pyrolysis temperatures, while later on the worms appeared to be more attracted to
soils with biochar produced at the highest pyrolysis temperature.
Partly supporting our results, Li et al. (2011) reported no effects of
10 g/kg of biochar on E. fetida, but the worms appeared to avoid the
same type of biochar at much higher rates (100 g/kg) unless the
biochar was wetted to its saturation point. The authors concluded
that drying of the soil due to the high rate of biochar was the main
cause for earthworm avoidance behavior. There is some evidence
suggesting that earthworms prefer, albeit only slightly, soils that
are amended with biochar to those without biochar (Chan et al.,
2008; Van Zwieten et al., 2010b). The short-term avoidance
behavior of earthworms in the current study may be due to slight
changes in the soil water potential, despite attempts to adjust soil
moisture separately in each pot according to the WHC of the soilbiochar mixtures.
Few data are available on biocharnematode interactions. Our
study showed that none of the biochar type exerted clear effects on
the total nematode abundance, suggesting that birch-produced
biochar, irrespective of pyrolysis temperature, is nontoxic to soil
inhabiting nematodes. Supporting our results, Zhang et al. (2013)
found no signicant effects of wheat straw biochar (400  C) on total
nematode abundance in stagnic anthrosols. Recently, reduced
number of root-knot and root-lesion nematodes in biochar treated
soils has been reported (George et al., 2016; Rahman et al., 2014).

232

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

However, as George et al. (2016) conclude, not all biochar types


may be suitable for nematode pest management, as also reported
for different types of compost (Akhtar and Alam, 1993). Differences
between the results of previously mentioned studies and our study
may thus be due to differences in biochar and soil types. It is also
worth mentioning that the conditions were not favorable for plant
feeding nematodes in our experiment as shown by the decreased
relative proportion of those nematodes during the experiment. The
numbers of herbivores decreased in all of the pots during the
incubation period probably because of the lack of host plants. In
addition, the rst test plant, radish, did not increase the abundance
of herbivores although numbers of bacterivores increased in the
biochar treated pots. It is well known that Brassicaceae crops can
be used to control plant parasitic species in cropping systems
(Fourie et al., 2016). Thus the radish may have had some
nematicidal impacts on herbivores. Consequently, the effects of
birch biochar on plant feeding nematodes may have remained non
detectable.
5.3. Plant yield
Our hypothesis that pyrolysis temperature determines the
effects of biochar on plant growth was partly supported: the
impacts of biochar on plant growth were plant specic. Results
showed that birch-derived biochar, can signicantly improve the
root yield of radish at relatively low dosage levels (20 g/L). In the
case of radish leaves, slight differences occurred between biochar
types, BC300 producing lower yield than BC375 or BC475.
However, none of the biochar types had an effect on the biomass
production of barley a plant species commonly used in biochar
tests. This is in contrast with Nelissen et al. (2014b) who showed
that the yields of radish and barley decreased in biochar-treated
soils; the decline increased with increasing pyrolysis temperatures. The authors attributed this to the very low NO3 availability
in biochar-treated soils, even in fertilized soils. The contrasting
results between our study and those by Nelissen et al. (2014b) are
supposedly due to differences in feedstock materials and pyrolysis
processes. Nelissen et al. (2014b) explored pine and willow biochar
pyrolysed at higher temperatures (450650  C) and the pyrolysis
time (3388 min) was shorter than in our study.
The cumulative yield of ryegrass, also a widely used test plant in
biochar studies, increased in all biochar-treated soils at higher
application rate (80 g/L). At the rst sampling event, all biochar
types reduced plant growth but later on the biochar, irrespective of
treatment, increased ryegrass growth. Differences in ryegrass
growth between the control and biochar-treated soils increased
during our study. Several harvests without the addition of fertilizer
are supposed to cause nutrient deciency in plants when nutrients
are removed with the harvested biomass. Increased ryegrass
growth in the biochar-treated soils during our study may be a
result of the release of biochar-adsorbed nutrients to the plants
(Taghizadeh-Toosi et al., 2012) or by enhanced microbial nutrient
mobilization in biochar treated soils (Fox et al., 2014). There is
evidence that mycorrhizal fungi growing on the surface of biochar
can induce phosphorus translocation from biochar to host roots
(Hammer et al., 2014). However, more knowledge on the indirect
effects of biochar on plant growth is needed. In addition, the slight
but statistically insignicant increase in the long-term soil water
content and the simultaneous increase in the yield of ryegrass in
biochar treated soils supports the idea of the ability of biochar to
improve water supply to plants (Cao et al., 2014; Kammann et al.,
2011). As hypothesized, there were some differences between the
three biochar types pyrolysed at different temperatures: compared
to BC375 and BC475, BC300 produced less ryegrass and radish
leave biomass. The reason for this remains unclear. Previously Liu
et al. (2013) showed in their meta-analysis study that biochar

pyrolysed at <350  C had no effect on crop productivity. According


to our results, biochar produced at 300 C (i.e. at lower temperature
than suggested by European Standard), also increases the growth
of radish and ryegrass, yet less than biochars produced at higher
temperatures (375475 C) at least in acidic sandy soil. As the
response rate of plants to various environmental factors tend to be
higher under laboratory conditions compared to eld conditions
(Liu et al., 2013), it is possible that the results of the current study
do not directly apply to eld conditions.
Complex chemical and/or soil-specic sorption processes
between biochar and the soil evidently affect the availability of
water and nutrients for plants and soil organisms. The application
of biochar in elds is not recommended in the spring before
cultivation, since biochar may absorb water and nutrients and
cause plant desiccation (Bruun et al., 2012; Novak et al., 2010;
Tammeorg, 2014). When applied in the autumn, biochar has the
potential to adsorb large quantities of water. Furthermore, water
and soluble nutrients stored in biochar during the fall may become
available for plant uptake in the following spring. These hypothesis
needs to be evaluated in the future.
5.4. Nitrogen concentration of plants
It is commonly reported that biochar may impact nitrogen
concentrations of plants (Clough et al., 2013). We hypothesized
that pyrolysis temperatures have an impact on the quality of
biochar in terms of affecting nutrient availability for plant uptake.
Nelissen et al. (2014b) observed that biochar pyrolyzed at high
temperatures (>450  C) can reduce plant N concentrations even at
low application levels (10 g/L). In their study, the decrease in
nitrogen became more pronounced with increasing pyrolysis
temperatures and was feedstock dependent, being larger for
willow than for pine biochar (Nelissen et al., 2014b). In our study,
birch-produced biochar at <475  C and applied at 20 g/L had no
effect on N content of lettuce. Instead, BC375 and BC475 reduced N
content of radish seedlings even at lower application rate (20 g/L).
At the higher application rate (80 g/L) the results were inconsistent
between plant species: BC300 had the negative effect on the N
content of lettuce while BC375 and BC475 affected radish seedlings
negatively. At the same time (results: Week 5), microbial activity
increased in biochar-treated (BC300 and BC375) soils indicating
that some of the N was incorporated into the microbial biomass
and was thus not available to plants. This temporary N immobilization after biochar application has been noted in several studies
and may explain part of our results (Bruun et al., 2012; Novak et al.,
2010; Tammeorg, 2014). The inconsistent results between plant
species in our study may be explained by differences in
experimental conditions. The lettuce experiment was performed
immediately after biochar application when biochar-induced
pyhytotoxicity (see above) was at a maximum and might also
have impacted lettuce metabolism. Instead, the radish experiment
was performed ve weeks after the stabilization period when most
phytotoxic compounds are likely to have degraded or volatilized.
Interestingly, even though radish seedlings growing in biocharamended soils (BC375, BC475) had lower N content than seedlings
growing in the control soil, all biochar types increased biomass
production and reduced the discoloration of radish. This was, to
some extent, an unexpected result since the lack of nitrogen and
the concomitant decrease in shoot N-content are generally
assumed to lead to a reduction in biomass production. The lush
growth of radish is usually associated with high availability of N in
the soil (Guvench, 2002). Evidently, nitrogen did not limit radish
growth in our study. The negative relationship between biochar
and radish tissue N concentrations could be explained by the
dilution effect whereby rapid plant growth can cause a small
decrease in nutrient concentrations of the plant (Jarrell and

M. Hagner et al. / Soil & Tillage Research 163 (2016) 224234

Beverly, 1981). However, it is not possible to distinguish between


effects caused by the immobilization of N by soil microbes, the
adsorption of N to biochar or the dilution effect. The reasons for the
reduced discoloration of radish seedlings in BC375 and BC475
treated soils remain unsolved. Biochar has been observed to reduce
and delay the onset of plant diseases (Elad et al., 2010; Ok et al.,
2016) which may partially explain the reduced discoloration in the
current study.
6. Conclusions
To summarize, in this study the effects of birch wood biochar,
produced at various temperatures, were investigated simultaneously on the many compartments of the soil ecosystem. Even
though the physical properties of the tested biochar types varied
considerably, all tested biochar types had parallel effects on soil
properties and plant growth. The differences that existed were
mostly slight and short-term. Our results showed that the effects of
biochar in soils are complex and context dependent. Consequently,
provision of accurate quality standards and guidelines for the
production and use of wood-based biochar is challenging. Practical
realism and more research are needed to enable the commercialization of wood-based biochar in agriculture.
Conict of interest
The authors declare that they have no conict of interest.
Compliance with ethical standards
The manuscript complies ethical rules applicable of this journal.
Acknowledgements
The authors thank the Finnish Funding Agency for Technology
and Innovation (Tekes) and several enterprises (e.g. Raussi Ltd.,
Charcoal Finland Oy and Biolan Oy) for funding this project. We
also thank the University of Helsinki, VTT Technical Research
Centre of Finland and Natural Resources Institute Finland (Luke)
personnel who participated in the research. Johan Kotze is thanked
for checking the English of the manuscript.
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