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ARTICLE IN PRESSG ModelGEE-4584; No. of Pages 9

Agriculture, Ecosystems and Environment xxx (2014) xxx–xxx

Contents lists available at ScienceDirect

Agriculture, Ecosystems and Environment

j ourna l h om epage: www.elsev ier .com/ locate /agee

hort-term effects of biochar on soil properties and wheat yieldormation with meat bone meal and inorganic fertiliser on a borealoamy sand

riit Tammeorga,∗, Asko Simojokib, Pirjo Mäkeläa, Frederick L. Stoddarda,aura Alakukkuc, Juha Heleniusa

Department of Agricultural Sciences, P.O. Box 27 (Latokartanonkaari 5, Plant Production Sciences), FIN-00014 University of Helsinki, FinlandDepartment of Food and Environmental Sciences, P.O. Box 27 (Latokartanonkaari 11, Environmental Soil Science), FIN-00014 University of Helsinki,inlandDepartment of Agricultural Sciences, P.O. Box 28 (Koetilantie 5, Agrotechnology), FIN-00014 University of Helsinki, Finland

r t i c l e i n f o

rticle history:vailable online xxx

eywords:iocharield experimentrganic fertiliserriticum aestivumater retention

ield components

a b s t r a c t

Poor water retention capacity (WRC) and nutrient deficiency commonly limit crop yields in sandy soils.The use of biochar as a soil amendment has been previously reported to improve these limiting factors insubtropical and temperate soils. We studied the effects of biochar on soil properties and yield formationof spring wheat (Triticum aestivum L.) when applied together with inorganic fertiliser or meat bone meal(MBM) to an Endogleyic Umbrisol with a loamy sand texture in boreal conditions. In a two-year fieldexperiment, biochar was applied at 0, 5, 10, 20 and 30 t ha−1 combined with three fertiliser treatments(unfertilised control, MBM and inorganic fertiliser) providing equal amounts of nitrogen (N), phosphorus(P) and potassium (K). Soil WRC and fertility as well as wheat yield, yield components and quality wereanalysed. Soil moisture content, leaf area index and leaf chlorophyll values (SPAD) were monitored duringthe experiment. Biochar increased the plant-available water content of the topsoil in the first year and

reduced the bulk density in the second year after application. It also increased the contents of easilysoluble K and soil organic C (SOC) in the 20 cm of topsoil, but had no effects on other soil nutrients, pHor moisture content. Biochar amendment decreased the soil NO3

−-N content below control values in thefirst year but increased it significantly in the second year. The addition of biochar did not significantlyaffect the nitrogen uptake, grain yield or quality of wheat, possibly because of its low nutrient availabilityand the high organic matter content of the soil.

. Introduction

The combination of biomass pyrolysis and use of the resultingiochar as a soil amendment simultaneously provides bioenergy,arbon (C) sequestration and soil conditioning (Lehmann et al.,008; Verheijen et al., 2009; Woolf et al., 2010). The long-term Cequestration potential of such biochar practices relies on the recal-itrance of the biochar-C to microbial decomposition (Lehmann

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t al., 2008; Singh et al., 2012). The improvement of soil fertilitys considered a further benefit of biochar application to acid orutrient-deficient (sub-) tropical soils (Steiner et al., 2008; Major

∗ Corresponding author. Tel.: +358 504 480 431.E-mail addresses: priit.tammeorg@helsinki.fi (P. Tammeorg),

sko.simojoki@helsinki.fi (A. Simojoki), pirjo.makela@helsinki.fi (P. Mäkelä),rederick.stoddard@helsinki.fi (F.L. Stoddard), laura.alakukku@helsinki.fiL. Alakukku), juha.helenius@helsinki.fi (J. Helenius).

167-8809/$ – see front matter © 2014 Elsevier B.V. All rights reserved.ttp://dx.doi.org/10.1016/j.agee.2014.01.007

© 2014 Elsevier B.V. All rights reserved.

et al., 2010; Vaccari et al., 2011; Zhang et al., 2012). In additionto the liming effect of certain biochars (Major et al., 2010; VanZwieten et al., 2010; Vaccari et al., 2011), the enhancement of soilfertility has been attributed to increased cation exchange capac-ity (Liang et al., 2006), fertilisation by nutrients contained in thebiochar (Major et al., 2010; Quilliam et al., 2012; Xu et al., 2013)and enhanced arbuscular mycorrhizal (AM) colonisation leadingto increased nutrient availability (Blackwell et al., 2010; Solaimanet al., 2010). Furthermore, the improved water retention capac-ity (WRC) of the soil (Eastman, 2011; Liu et al., 2012) may explainsome of the previously reported increases in the activity of soil biota(Chan et al., 2008; Lehmann et al., 2011), enhanced nutrient use effi-ciency (Chan et al., 2007; Steiner et al., 2008) and crop yields (Majoret al., 2010; Vaccari et al., 2011; Zhang et al., 2012) on subtropical

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

soils.The effects of biochar on soil and plant properties vary widely,

depending on the characteristics of both the underlying soil andthe biochar. This is demonstrated by the lack of significant changes

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ash. It comprised 883 g total C kg−1, 4.66 g Ca kg−1, 4.52 g K kg−1 and3.5 g N kg−1. Additional details about the physicochemical proper-ties of the biochar are presented in Tammeorg et al. (2014b).

Table 1Mean air temperature (◦C) and precipitation (mm) in Helsinki for growing seasons2011–2012 and the long-term (1971–2000) averages at Helsinki Kaisaniemi (FMI,2012, 2013).

Month Mean temperature (◦C) Precipitation (mm)

1971–2000 2011 2012 1971–2000 2011 2012

May 9.9 9.9 10.9 32 27 65June 14.8 16.7 13.7 49 49 88

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n crop yields 1–4 years after biochar application to fertile min-ral soils in temperate (Güerena et al., 2012; Jones et al., 2012)nd boreal climates (Karhu et al., 2011; Tammeorg et al., 2014a),n spite of improvements in nutrient and water status of the soils.he challenges in scaling up the now rather sporadic use of biochars a soil amendment relate largely to the uncertainties concern-ng its long-term effects on soil quality, as it cannot practically beemoved from soil after application (Jones et al., 2012). The pos-ible negative effects of biochar soil addition include short-termeductions in mineral nitrogen (N) availability in soils (Novak et al.,010; Bruun et al., 2012; Tammeorg et al., 2012), decreased perfor-ance of crops on calcareous soils (Kishimoto and Sugiura, 1985;an Zwieten et al., 2010), increased loss of native soil C (Wardlet al., 2008; Zimmerman et al., 2011), and decreased availability ofoil-applied herbicides (Graber et al., 2012). Furthermore, little isnown about the biochar-mediated changes on soil and plant prop-rties in boreal soils affected by freeze-thaw cycles (Karhu et al.,011), and considering the high availability of different biocharaw materials in northern countries (e.g., forestry residues), theres an urgent need for scientific evidence on both the positive andegative long-term effects of biochar on the soil-plant-atmosphereystem in this region.

A few studies have been published on the interactive effectsf biochar and organic fertilisers applied together (Lehmann et al.,003; Steiner et al., 2007, 2008; Schulz and Glaser, 2012; Tammeorgt al., 2012). The importance of nutrient recycling through ampli-ed use of organic fertilisers is increasingly acknowledged (Royt al., 2002; Römer, 2009; Fischer and Glaser, 2012) as the inputs fornorganic fertiliser production (e.g., phosphorus (P) rocks) becomeepleted (Cordell et al., 2009) and the prices of inorganic fertilis-rs increase (Silva, 2011; USDA, 2013). The interactive mechanismsetween biochar and organic fertilisers have been associated with

ncreased contents of SOC and black carbon that could enhancehe nutrient retention capacity of the soil (via increased cationxchange capacity and forming of organo-mineral complexes;laser et al., 2002) and improve the sorption capacity of phytotoxicubstances (Hille and Den Ouden, 2005; Schulz and Glaser, 2012).

greater increase in biomass of oat (Avena sativa L.) was reportedrom a biochar-compost mixture than from pure compost addi-ion under tropical conditions (Schulz and Glaser, 2012). In borealonditions, the effects of soil-applied biochar on N mineralisationynamics depended greatly on the C:N ratios of the organic fer-ilisers (Tammeorg et al., 2012). Understanding the effects of usingiochar together with organic fertilisers is particularly importanthen biochar is used as a soil amendment in organic farming sys-

ems.One of the organic fertilisers at present increasingly used in

urope is a co-product of the meat processing industry, meat boneeal (MBM), which has a low C:N ratio (about 4.5) facilitating faster

mineralisation than many other organic fertilisers (Salomonssont al., 1994; Tammeorg et al., 2012). Similarly to biochar, it haseen associated with enhanced activity of soil microbes (Mondinit al., 2008), increments in the N and P use efficiencies of cropsYlivainio et al., 2008; Jeng and Vagstadt, 2009) and improved cropields (Jeng et al., 2004, 2006; Chen et al., 2011). Thus, if MBM ispplied together with biochar, it is possible that certain interactiveechanisms leading to agronomic benefits may follow.It has been suggested that the effects of biochar in temperate

oils would be most evident in coarse-textured soils by mediationf their nutrient deficiency and poor WRC (Atkinson et al., 2010;iu et al., 2012). In our previous laboratory study (Tammeorg et al.,012), the application of biochar together with MBM to a loamy

Please cite this article in press as: Tammeorg, P., et al., Shyield formation with meat bone meal and inorganic fertiliser

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and in boreal conditions facilitated initial N immobilisation in aose-dependent manner. To gain further insights into the mech-nisms of the effects of biochar in combination with organic andnorganic fertiliser treatments on the physicochemical properties

PRESS and Environment xxx (2014) xxx–xxx

of soil and crop performance on a field scale, a field experiment withfive biochar application rates was started on the same nutrient-deficient, coarse-textured soil as used in the laboratory study. Thespecific aims of this study were to determine the short-term effectsof different biochar application rates on (i) the physicochemicalproperties of soil; (ii) the growth dynamics, N uptake (NU), yieldand quality of spring wheat (Triticum aestivum L. emend Thell.), aswell as (iii) to evaluate whether the effects of biochar applicationon wheat growth and soil properties depend on the type of thefertiliser.

2. Materials and methods

2.1. Experimental site and soil

The field experiment was conducted over two consecutivegrowing seasons (2011 and 2012) at the Viikki Experimental Farm,University of Helsinki, Finland (60◦13′42′′ N 25◦2′34′′ E). For thepreceding six years, the field was cropped with spring wheat andbarley (Hordeum vulgare L.) with conventional mouldboard plough-ing to 20–25 cm depth and inorganic fertiliser application. Thesoil was classified as an Endogleyic Umbrisol (WRB, 2007) with aloamy sand texture with 83% sand, 15% silt and 2% clay (Soil SurveyDivision Staff, 1993) at 0–30 cm depth. The original content of soilorganic matter (SOM) was 63.4 g kg−1, assuming a 50% C contentfor the SOM (Pribyl, 2010). Before the start of the experiment, thesoil had a sufficient level of easily soluble P (21 g m−3 soil, extrac-tion with 0.5 M ammonium acetate, pH 4.65), whereas the levelsof easily soluble Ca, K, Mg and S were deficient according to theFinnish classification of arable soils (Viljavuuspalvelu Oy, 2008).

The growing season of 2011 was notably warmer than the 30-year average in Helsinki, especially in July and August (Table 1). Thetemperatures in 2012 were close to the long-term mean. August2011 and May, June and September 2012 were much wetter thanthe long-term means.

2.2. Biochar

The biochar was obtained by pyrolising chips of debarked spruce(Picea abies (L.) H. Karst.) in a continuously pressurised carboniser(Preseco Oy, Finland) at 550–600 ◦C. Air-dried chips were fed intothe reactor tube via an airtight system and subsequently movedby a screw conveyor through the hot region of the reactor tubein 10–15 min. The biochar was cooled overnight in an airtight siloand then ground in a roller mill. Before application to the soil, thebiochar was wetted to 25% (w/w) to moderate dust problems. Thebiochar had a pHH2O of 8.1, a specific surface area of 265 m2 g−1

and contained 122 g kg−1 of volatile matter (VM) and 27 g kg−1 of

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

July 17.2 20.6 17.7 62 56 54August 15.8 17.5 16.0 78 173 39September 10.9 13.6 12.5 66 88 160

Mean 13.7 15.7 14.2 Sum 287 393 406

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Table 2The experimental treatments and their coding.

Biochar addition(B; t ha−1)

Fertilisation (F) Mean

Unfertilisedcontrol

Meat-bone-meal

Inorganicfertiliser

0 B0F0 B0FM B0FI B0

5 B5F0 B5FM B5FI B5

10 B10F0 B10FM B10FI B10

20 B20F0 B20FM B20FI B20

30 B30F0 B30FM B30FI B30

Tp

2

istasebrafs4

fbatb(aiaHctN

cCTfuwtFe2cimf

aawBmU

Mean F0 FM FI

reatment combinations chosen for measurements of soil moisture and soil physicalroperties are shown in bold.

.3. Experimental setup

The field experiment was established in May 2011 by apply-ng biochar to the experimental plots (2.2 m × 10 m) with a sandpreader and mixing it into the uppermost 10 cm of the soil bywo opposite passes of a rotary power harrow. The biochar waspplied only once during the experiment and buffer plots of theame size as the experimental plots were used between the differ-nt biochar treatments to minimise cross-contamination of plotsy wind-blown biochar dust. The buffer plots and the control plotseceived the same tillage treatment as the biochar plots. The dayfter biochar application, spring wheat cv ‘Amaretto’ was sown andertilised with a combine seeder at 650 viable seeds m−2 and rowpacing 12.5 cm. The sowing and fertiliser placement depths were–5 cm and 6–7 cm, respectively.

The experiment was set up with a factorial split-plot design withour complete blocks as replicates. The main plot factor was theiochar application rate (B) (0, 5, 10, 20 and 30 t dry matter ha−1)nd the sub-plot factor was the fertiliser treatment (F) (Table 2). Thereatment combinations were coded as BXFY, where X refers to theiochar application level (t ha−1) and Y to the fertiliser treatment0, M and I corresponding to unfertilised control, meat bone mealnd inorganic fertiliser, respectively). The inorganic fertiliser usedn the experiment was Agro 28-3-5 (Cemagro Oy, Lohja, Finland)nd the MBM-based fertiliser was Aito-Viljo 8-5-2 (Honkajoki Oy,onkajoki, Finland), where the numbers refer to the elementalontents of N, P and potassium (K) (w/w %), correspondingly. Inhe inorganic fertiliser, the N was in the form of both NO3

− andH4

+.As the plant availability of N in MBM has been reported to be

omparable with that of inorganic N fertilisers (Jeng et al., 2004;hen et al., 2011), both fertilisers were applied at 100 kg N ha−1.his amount delivered comparable amounts of plant-available Por the first year, assuming 18% of total MBM-P being water sol-ble in the first growing season (Ylivainio and Turtola, 2009). Itas expected that 7% of the P from MBM would be released during

he second growing season (Ylivainio and Turtola, 2009), so Yaraosforiravinne® (9% P) was added to the FI treatment in 2012 toqualise the P fertilisation. The applied amount of MBM delivered

kg more K per hectare than the inorganic fertiliser, so K2SO4 (Kontent 41.5%, Yara Suomi Oy, Siilinjärvi, Finland) was added to thenorganic fertiliser treatment. Thus, in 2011, both fertiliser treat-

ents delivered 10.8 kg P ha−1 and 19.5 kg K ha−1 in easily solubleorm, and in 2012, 14 kg P ha−1 and the same amount of K.

Integrated management practices were used, including thepplication of chemical herbicides, fungicides and pesticides acrossll treatments when appropriate. In both years, at tillering, the crop

®

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as sprayed with growth regulator chlormequat (Cycocel 750,ASF Agro, Ludwigshafen, Germany) at 0.8 L ha−1, and the herbicideixture of mecoprop-P, MCPA and dichlorprop-P (K-Trio® NufarmK Ltd, Kent, UK) at 1.2 L ha−1, and at heading with the fungicide

PRESS and Environment xxx (2014) xxx–xxx 3

pyraclostrobin (Comet®, BASF Agro, Limburgerhof, Germany)at 0.3 L ha−1. Additionally, the insecticide alpha-cypermethrin(Fastac®, BASF Agro, Limburgerhof, Germany) and the fungicidemixture of tebuconazole and protioconazole (Prosaro®, BayerCropScience AG, Monheim am Rhein, Germany) were both appliedat 0.3 L ha−1 at heading in 2011, while the insecticide-acaricidetau-fluvalinate (Mavrik®, Yates, New Zealand) was applied attillering in 2012 at 0.2 L ha−1. After the first growing season, thefield was disc-harrowed to 12 cm, and was rotary power harrowedto 10 cm before sowing and fertilisation of wheat in spring 2012.

2.4. Sampling, measurements and analyses

For chemical analyses, 10 soil samples were taken from the top20 cm (core diameter 2.7 cm) and mixed to form a composite sam-ple from each experimental plot prior to starting the experimentand in the autumns of 2011 and 2012. Soil chemical analyses wereconducted by an acid (pH 4.65) ammonium acetate extraction on avolumetric basis (Vuorinen and Mäkitie, 1955). The extraction wasfollowed by the elemental analysis of the extracts by inductivelycoupled plasma optical emission spectrometry (ICP-OES; Thermo-Fisher iCAP6500, Thermo Fisher Scientific, Cambridge, UK), withthe exception of P that was determined by colorimetry with amolybdenum blue method (Lachat QuikChem 8000, Lachat Instru-ments, Milwaukee, USA). Electrical conductivity and pH of soil weremeasured from a 1:2.5 (w/w) soil-to-water mixture (Vuorinen andMäkitie, 1955). The contents of soil organic C (SOC) and total Nin soil were measured with a VarioMax CN analyser (ElementarAnalysensysteme GmbH, Germany). Soil C was considered to beorganic since the carbonate content of this soil was known to benegligible. The mineral N (NH4

+-N and NO3−-N) content of all

soil samples was extracted from soil samples with 2 M KCl solu-tion (1:5 v/v) for 1 h. The ammonium and nitrate concentrationsof the extracts were determined spectrophotometrically with anFIAstar 5000 Flow Injection Analyzer (Foss Tecator AB, Höganäs,Sweden). The mineral N analyses of all soil samples were con-ducted in autumn 2012. The samples were stored at −20 ◦C for 3–19months before this.

Soil moisture content was measured weekly in B0F0, B10F0,B30F0, B0FI, B10FI, B30FI treatments using time domain reflectrom-etry (TDR, MiniTrase 6050X3, Soilmoisture Equipment, USA) atdepths of 0–15, 0–28 and 0–58 cm. The soil moisture content wasmeasured at one point at each depth and the contents in layers15–28 cm and 28–58 cm were calculated. Weekly mean and max-imum air temperatures and the precipitation data were acquiredfrom the Vaisala WXT520 automatic weather transmitter (VaisalaOy, Vantaa, Finland) 1.6 km from the field.

The water retention at matric suctions 3–1500 kPa (pF 1.5–4.2)of topsoil (2.5–7.5 cm) was measured from the same six treatments(Table 2) at the end of both growing seasons by taking four undis-turbed soil samples from each treatment plot into 100 cm3 steelcylinders. The WRC of the samples was determined with the sand-box method at matric suctions 3 and 6 kPa and with the pressureplate method at matric suctions 10, 50, 250 and 1500 kPa (Daneand Hopmans, 2002). Additionally, the same samples were usedto calculate the plant available water content (AWC; the differencebetween the water contents at 6 kPa and 1500 kPa) and the dry bulkdensity. The total porosity was calculated from the bulk densityassuming a density of 2.65 g cm−3 for soil particles.

Plant stand density was determined by counting the number ofplants in three representative lengths of 30 cm of the sowing row

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

at the leaf development growth stage (GS 12; BBCH, Meier, 2001).Every week from stem elongation to grain filling, the relative leafchlorophyll values (SPAD) of the uppermost fully developed leafwere acquired from 20 plants per plot with a SPAD-502 portable

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hlorophyll meter (Minolta Camera Co. Ltd., Osaka, Japan) and leafrea index (LAI) was measured with a SunScan SS1 ceptometerar (Delta-T Devices Ltd., Cambridge, UK) four times in each plot.bove-ground plant biomass (AGB) was sampled at GS 29, 65 and85 (BBCH, Meier, 2001). The AGB was sampled within a 2 m × 2 mrea near one end of the plot by cutting the plants at 2 cm abovehe soil surface from three randomly chosen 30 cm row lengths. Thelant samples were dried in an oven for 72 h at 60 ◦C and weighed.he first two samplings were ground by hammer mill (screen size

mm; Koneteollisuus Oy, Helsinki, Finland) and analysed for C and content with the VarioMax CN analyser. The nitrogen uptake (NU)t anthesis (GS 65) was calculated by multiplying the AGB by the Nontent. The number of plants in the third growth stage sample wasecorded and plants were divided into vegetative mass (leaves andtem), and spikes, which were counted. Samples were threshed,rain number and weight were recorded, and harvest index (HI)alculated.

Grain was harvested from an area of 11.25 m2 (1.5 m × 7.5 m)rom the middle of a plot by a combine harvester, dried, sortednd weighed, and a 500-g subsample analysed for quality. Theoisture content was determined by weighing the water loss of

0 g of grains dried for 16–18 h in an oven at 105 ◦C and the 1000rain weight (TGW) was determined. The starch and protein con-entrations were determined with a Perten DA 7200 near infraredpectroscope (NIR) (Perten Instruments, Huddinge, Sweden).

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.5. Statistical analyses

The effects of biochar and fertiliser on the changes in soil chemi-al properties from the initial conditions (spring 2011) were tested

able 3he change in soil chemical composition in the 0–20 cm layer in autumns 2011 and 2012011), together with the probability values for treatment factors and interactions. Data sh

Year Treatment EC pH Acid ammonium acetat

(�S cm−1) Ca P K

2011 spring Mean 75.8 6.35 1127 20.6 6

2011 B0 −11.7 −0.06 ab 75 b 0.7 5B5 −15.8 −0.18 a −48 a −0.6 3B10 −14.2 −0.03 ab 11 ab 1.3 0B20 −15.0 0.03 b 17 ab 1.5 1B30 −20.0 0.04 b 12 ab 2.1 2SEM 3.3 0.03 18 0.9 2

F0 −19.0 −0.03 8 ab −1.5 a 1FM −12.0 −0.01 49 b 5.4 b 1FI −15.0 −0.08 −16 a −1.0 a 1SEM 3.3 0.05 19 1.0 2

df

B 4 n.s. 0.004 0.043 n.s. <F 2 n.s. n.s. 0.013 <0.001 nB × F 8 n.s. n.s. n.s. n.s. n

2012 B0 −12.5 −0.09 ab 15 b 0.6 −B5 −14.2 −0.18 a −90 a 0.2 −B10 −20.8 −0.10 ab −32 ab 0.1 −B20 −21.7 −0.04 b −58 ab 0.7 1B30 −11.7 −0.06 b −17 ab 1.7 1SEM 3.6 0.02 16 0.5 2

F0 −21.0 −0.05 b −38 −1.8 a 3FM −14.5 −0.08 b −12 5.1 b −FI −13.0 −0.16 a −60 −1.3 a −SEM 3.7 0.02 21 0.7 2

df

B 4 n.s. 0.008 0.036 n.s. 0F 2 n.s. 0.001 n.s. <0.001 0B × F 8 n.s. n.s. n.s. n.s. n

ositive means represent additions and negative values show decreases from the springetween treatment means within single years according to Tukey’s HSD test. The significareatment; EC = Electrical conductivity; Nmin = (NH4

+-N + NO3−-N); Corg = soil organic C; F0

rror of the means for all the respective biochar or fertiliser treatments; df = degrees of fr

PRESS and Environment xxx (2014) xxx–xxx

with two-way split-plot analysis of variance (ANOVA) with biocharlevel, fertiliser type, and their interactions as fixed effects. Meanswere compared with the Tukey HSD multiple pair-wise compari-son test. The effects of treatment on soil physical properties andplant properties were tested with two-way analysis of covariance(ANCOVA), using the underlying highly variable soil C content as thecovariate, and comparing the adjusted least-square means. Bonfer-roni correction was used in the post hoc tests of the ANCOVA formultiple comparisons, which resulted in different standard errorsfor each difference between means. Additionally, Pearson corre-lation coefficients were estimated between biochar applicationrates, chemical properties of soil and parameters of wheat growthand yield. Statistical analyses were conducted with the softwarepackage PASW v 21.0 (SPSS Corp., Chicago, USA) using p < 0.05 asthe threshold for significance.

3. Results

3.1. Soil properties

Increasing quantities of biochar increased the SOC contents inautumn 2011, with the B20 and B30 treatments being significantlyhigher than the control, but the differences were no longer signif-icant in 2012 (Table 3). Similarly, the increased quantity of easily

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

soluble K of the soil in both years over the control (positive correla-tion between biochar rates and K content; Supplementary Table A1)was significant only at the highest application rates (B20 and B30 in2011 and B30 in 2012). The highest biochar addition decreased the

compared to the corresponding properties at the beginning of experiment (springow means of 4 replicates across 3 fertilisation treatments or 5 biochar levels.

e extractable (g m−3 soil) 2 M KCl-extractable (g m−3 soil) Total (g kg−1)

Mg S NH4+-N NO3

−-N Nmin N Corg

2 100 5.2 6.2 5.5 11.7 2.4 31.7

a 10 b 1.2 −0.4 0.2 b −0.2 ab −0.2 −1.0 a a −2 a 0.9 −0.4 −0.1 b −0.4 ab −0.2 1.6 ab

a 6 ab 1.3 −1.6 −0.5 ab −2.2 ab −0.2 0.8 ab7 b 7 ab 1.5 1.0 −0.2 b 0.8 b −0.2 2.9 bc5 b 7 ab 1.5 −1.2 −1.6 a −2.8 a −0.1 5.0 c

2 0.3 0.5 0.2 0.5 0.1 0.9

0 7 1.6 b −0.5 −2.0a −2.5 a −0.3 1.80 7 1.7 b −0.3 0.2b −0.1 b −0.2 1.70 3 0.5 a −0.8 0.5b −0.2 b −0.1 2.1

2 0.3 0.7 0.3 0.8 0.1 0.7

P-values0.001 0.044 n.s. 0.083 0.011 0.009 n.s. 0.027.s. n.s. 0.002 n.s. <0.001 0.023 n.s. n.s..s. n.s. n.s. n.s. n.s. n.s. n.s. n.s.

7 a −3 2.0 −1.1 −1.3 a −2.3 0.2 0.27 a −12 1.6 −0.7 −0.3 ab −1.0 0.0 −0.65 a −7 1.0 −0.4 −0.9 ab −1.3 0.1 1.9

a −7 0.8 0.0 −0.4 ab −0.4 0.1 1.71 b −6 2.3 −1.1 0.3 b −0.8 0.0 2.5

1 0.6 0.3 0.2 0.4 0.1 1.1

b −6 0.8 −0.7 −1.7 a −2.4 0.1 1.24 a −7 0.8 −0.4 0.0 b −0.4 0.1 1.53 ab −8 3.0 −0.9 0.1 b −0.7 0.1 0.8

3 0.9 0.7 0.3 0.8 0.1 1.3

P-values.002 0.026 n.s. n.s. 0.008 n.s. n.s. n.s..015 n.s. 0.050 n.s. <0.001 0.051 n.s. n.s..s. n.s. n.s. n.s. n.s. n.s. 0.087 0.086

2011 values. Different lowercase letters indicate significant differences (p < 0.05)nt p values (p < 0.05) are bolded. Abbreviations: B = biochar treatment; F = Fertiliser= unfertilised control, FM = meat bone meal; FI = inorganic fertiliser; SEM = standard

eedom; n.s. = not significant (p > 0.1).

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oil NO3−-N content from control values in 2011, but in the autumn

012, the effect was the opposite (Table 3).In both years, the B30 treatments increased the soil pH over the

5 treatments (0.2 and 0.1 units for 2011 and 2012, respectively),ut the differences from the control were not significant. The B5reatment resulted in a significant decrease in easily soluble Cand Mg below control values in both years, whereas there wereo significant differences between the control and higher biocharpplication levels (Table 3).

In both 2011 and 2012, the MBM treatments increased the eas-ly soluble P content over that in inorganic fertiliser treatmentsnd unfertilised controls, whereas the amounts of easily soluble Cand S were significantly increased only in 2011 (Table 3). In bothears, both the fertilisers increased the soil NO3

−-N and mineral Nontents over the control, but there were no differences betweenhe fertilisers. The biochar × fertiliser interaction was not signifi-ant in any of the soil chemical composition properties measuredn 2011–2012.

The B10 and B30 treatments had no significant (p > 0.1) effectsn the soil moisture content in comparison to the control at anyepth and any time, although the moisture content of the 0–15 cmoil layer in the B30 treatments was higher than that of the B10reatments at flowering and after harvesting in 2011 and from leafevelopment to crop maturity in 2012 (Fig. 1). The higher temper-tures in 2011 resulted in drier topsoil than in 2012. The moistureontent of the topsoil never decreased close to the PWP in 2012,

Please cite this article in press as: Tammeorg, P., et al., Shyield formation with meat bone meal and inorganic fertiliser

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hereas in 2011 this occurred during flowering and grain fillingFig. 1). The addition of inorganic fertiliser reduced the moistureontent of both 0–15 cm and 15–28 cm soil layers below unfer-ilised control values after the plants reached anthesis, whereas

ig. 1. The moisture content of topsoil (0–15 cm) as related to wheat growth stages, precipnd different lowercase letters indicate the significant differences between the Bonferronn a given week at p < 0.1 (+), p < 0.05 (*) and p < 0.01 (**). Note that the standard errors o012, respectively. PWP refers to the permanent wilting point moisture (1500 kPa; mean

PRESS and Environment xxx (2014) xxx–xxx 5

neither biochar nor fertiliser treatments had any significant effecton soil moisture content in the 28–58 cm layer (p > 0.05; data notshown). In the deepest layer, biochar reduced the decreasing effectof fertiliser addition on the soil moisture content before tillering in2012 (biochar × fertiliser interaction p < 0.05, data not shown).

The highest biochar addition decreased soil dry bulk density andincreased porosity over B10 treatments in the 2.5–7.5 cm layer in2011, but the differences between the control and biochar treat-ments were not significant (Table 4). In 2012, B30 treatments hadsignificantly lower soil bulk density and higher porosity than thecontrol, while the values of B10 treatments were not different fromthose of the other treatments. Similarly, the addition of inorganicfertiliser decreased soil bulk density and increased porosity in 2012.The B30 treatment increased the AWC and the WRC of the soil at6 kPa significantly over control values in 2011 but not significantlyin 2012 or at other matric potentials (Table 4, Supplementary Fig.A1). In 2011, the FI treatment decreased the soil WRC at 6 kPa fromthe F0 treatment when combined with B0 treatment, but in the B10and B30 treatments the opposite trend was seen (biochar × fertiliserinteraction significant, Table 4).

3.2. Plant properties

The wheat AGB, its N content and the NU as well as the SPADand LAI values increased generally in order of F0 < FM < FI in bothyears (Supplementary Table A2). Biochar application had no sig-

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

nificant effects on LAI, SPAD, AGB, N content and NU at tilleringor flowering (Supplementary Tables A1 and A2). Similarly, at othergrowth stages, the biochar level did not affect significantly the LAIand SPAD values in either year, but there was a trend of lower LAI

itation and the mean air temperature during growing seasons 2011–2012. Asterisksi-adjusted least-square means of the moisture content between biochar treatmentsf differences between the means ranged between 0.7–2.6 and 0.5–3.0 in 2011 and

of all treatments 2011 and 2012).

Please cite this article in press as: Tammeorg, P., et al., Short-term effects of biochar on soil properties and wheatyield formation with meat bone meal and inorganic fertiliser on a boreal loamy sand. Agric. Ecosyst. Environ. (2014),http://dx.doi.org/10.1016/j.agee.2014.01.007

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Table 4The physical properties of the topsoil (2.5–7.5 cm depth) in autumns 2011 and 2012 and probability values for treatment factors and interactions. Data show means of 4replicates across 2 fertilisation treatments or 3 biochar levels.

Year Treatment Bulk density Porosity Water content % by volume

(g cm−3) % 3 kPa 6 kPa 1500 kPa AWC

2011 B0 1.00 ab 62.3 ab 41.2 ab 27.3 a 7.3 20.1 aB10 1.11 b 58.2 a 42.2 a 30.0 ab 8.3 21.7 abB30 1.04 a 60.9 b 43.9 b 31.2 b 7.8 23.4 bSEM 0.01 0.4 0.4 0.4 0.3 0.4

F0 1.05 60.3 41.9 a 29.3 7.7 21.6FI 1.05 60.6 43.0 b 29.7 7.9 21.8SEM 0.01 0.4 0.3 0.2 0.2 0.2

df P-valuesB 2 0.010 0.010 0.043 0.038 n.s. 0.023F 1 n.s. n.s. 0.019 n.s. n.s. n.s.B × F 2 0.068 0.063 n.s. 0.014 n.s. n.s.

2012 B0 1.21 b 54.4 a 43.4 33.9 8.9 25.0B10 1.07 ab 59.5 ab 38.9 28.3 7.8 20.4B30 1.02 a 61.5 b 41.4 32.3 8.0 24.3SEM 0.02 0.7 0.6 1.1 0.4 0.8

F0 1.13 b 57.5 a 41.6 32.3 8.3 24.0FI 1.07 a 59.5 b 40.8 30.7 8.2 22.5SEM 0.01 0.5 0.6 0.6 0.2 0.5

df P-valuesB 2 0.029 0.026 n.s. n.s. n.s. 0.079F 1 0.035 0.037 n.s. n.s. n.s. 0.091B × F 2 n.s. n.s. n.s. n.s. n.s. n.s.

Different lowercase letters indicate significant differences (p < 0.05) between Bonferroni-adjusted least-square means within single years. Note that the differences betweenthe means had different standard errors. The significant p values (p < 0.05) are bolded. Abbreviations: B = biochar treatment; F = Fertiliser treatment; F0 = unfertilised control,FI = inorganic fertiliser; SEM = mean standard error of the means for all the respective biochar or fertiliser treatments; df = degrees of freedom; AWC = available water content.

Table 5The yield, yield components, and quality parameters of spring wheat (14% moisture content), with the probability values for treatment factors and interactions for the years2011 and 2012. Data show means of 4 replicates across 3 fertilisation treatments or 5 biochar levels.

Year Treatment Plants (m−2) Grains (plant−1) HI Yield (t ha−1) TGW (g) Starch (g kg−1) Protein (g kg−1)

2011 B0 558 24.1 0.48 3.8 32.2 70.4 10.1B5 485 24.9 0.46 3.5 33.1 70.3 10.3B10 469 28.1 0.45 3.6 33.0 70.2 10.4B20 519 23.3 0.46 3.4 31.8 70.2 10.1B30 508 28.2 0.47 4.1 33.7 70.6 9.9SEM 21 2.3 <0.01 0.2 0.3 0.1 0.2

F0 520 19.4 a 0.44 a 2.2 a 30.8 a 70.6 b 8.9 aFM 496 27.0 b 0.45 a 4.1 b 33.4 b 70.6 b 9.7 bFI 507 30.7 b 0.49 b 4.7 c 34.1 b 69.9 a 11.8 cSEM 27 1.5 <0.01 0.1 0.4 0.1 0.1

df P-valuesB 4 n.s. n.s. n.s. n.s. n.s. n.s. n.s.F 2 n.s. <0.001 <0.001 <0.001 <0.001 <0.001 <0.001B × F 8 n.s. n.s. 0.003 n.s. n.s. n.s. n.s.

2012 B0 382 27.4 0.29 2.0 23.7 70.2 11.2B5 341 29.8 0.31 1.9 25.1 70.2 11.4B10 331 33.7 0.32 1.8 24.9 70.0 11.4B20 344 26.4 0.30 1.6 24.1 70.1 11.2B30 349 31.2 0.31 1.9 24.3 70.1 11.6SEM 14 2.2 0.01 0.1 0.4 0.2 0.2

F0 335 17.7 a 0.27 a 0.9 a 22.2 a 70.1 a 11.3 bFM 350 33.6 b 0.31 b 1.9 b 24.1 b 70.4 b 10.8 aFI 363 37.7 b 0.34 c 2.7 c 27.0 c 69.9 a 12.0 cSEM 14 2.7 0.01 0.1 0.3 0.1 0.1

df P-valuesB 4 n.s. n.s. n.s. n.s. n.s. n.s. n.s.F 2 n.s. <0.001 <0.001 <0.001 <0.001 <0.001 <0.001B × F 8 n.s. n.s. n.s. n.s. n.s. 0.006 0.002

Lowercase letters indicate significant differences (p < 0.05) between Bonferroni-adjusted least-square means within single years. Note that the differences between the meanshad different standard errors. The significant p values (p < 0.05) are bolded. Abbreviations: B = biochar treatment; F = fertiliser treatment; F0 = unfertilised control, FM = meatbone meal; FI = inorganic fertiliser; HI = harvest index; TGW = 1000 grain weight; SEM = standard error of the means for all the respective biochar or fertiliser treatments;df = degrees of freedom.

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alues in B5 and B10 treatments than in the B0 and B30 treatmentsuring flowering in 2011 (p < 0.1; data not presented). The biochar xertiliser interaction was significant for only one of the determinedheat canopy growth properties, namely the higher N content of

he AGB in FI treatments over the FM treatments in the presence of10 and B20 at tillering in 2011 (Supplementary Table A2).

Biochar addition had no significant (p > 0.05) effect on the planttand density, grain number per plant, HI, grain yield or grain qual-ty (Table 5, Supplementary Table A1). The fertilisers increasedhe number of grains over the unfertilised control in both yearsy 39–113% (Table 5) and the difference was through increasedumber of grains per spike rather than increased number of spikeser plant (data not shown). The HI and grain yield were highest

n FI, followed by the FM and F0 treatments. In 2011, the fertiliser-elated increase in the HI was most pronounced in the B5 treatmentbiochar × fertiliser interaction significant). Grain yields were pos-tively correlated with the contents of soluble N and P and total Nn the first year and the contents of soluble N and S in the secondear (Supplementary Table A1).

The FI treatment was associated with higher TGW and grain pro-ein content than in F0, while the FM treatment resulted in valuesetween those of the other two (Table 5). In 2012, the B20 and30 treatments resulted in higher protein and lower starch con-ent of grains than the B0 treatments in unfertilised plots, but nouch effect was observed in the FM and FI plots (biochar x fertilisernteraction significant; data not shown).

. Discussion

.1. Physicochemical properties of soil

Application of biochar resulted in significant increases in SOCnly when the amount applied was sufficiently large (20 t ha−1) toe detected above background variation, as has been found pre-iously, where minimum biochar application rates of 10–25 t ha−1

Jones et al., 2012; Liu et al., 2012; Tammeorg et al., 2014a) wereeeded for increases in SOC content to be significant. In the sec-nd year after application, at the highest biochar application rate30 t ha−1, about 40% of the original SOC content of the 0–20 cmoil layer) only 18% of the biochar-added C was detected, resultingn no significant differences in the SOC content from the control.his decrease is consistent with previous field studies (Jones et al.,012; Tammeorg et al., 2014a) and is probably caused by downwardovements (Petter et al., 2012; Tammeorg et al., 2014a) of the fine

iochar particles to the subsoil layers of this coarse-textured soily earthworm activity, root growth and leaching.

The decreased soil NO3−-N content in the B30 treatment com-

ared with the control (Table 3), together with the decreasing trendn LAI values in the B5 and B10 application levels in 2011, may haveeen caused by initial N immobilisation. The biochar-mediatedhort-term N immobilisation has previously been reported bothn laboratory incubations (Novak et al., 2010; Bruun et al., 2012;ammeorg et al., 2012) and in the field (Lehmann et al., 2003;ammeorg et al., 2014a). This may be attributed to the small pro-ortion (0.1–0.27%) of labile C in biochar from woody raw materialshat decomposes within the first months after application dueo oxidation reactions of surface functional groups (Hamer et al.,004; Jones et al., 2011; Singh et al., 2012) and the high C:N ratioRajkovich et al., 2012) of the biochar used (251:1). In our previousaboratory study with the same soil (Tammeorg et al., 2012), weound a biochar dose-dependent immobilisation of N in unfertilisednd MBM treatments, whereas the reductions in mineral N started

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o decrease two months after application, possibly by the turnoverf microbial biomass. The highest NO3

−-N content of the soil in the30 treatments and the increased grain protein content comparedith the control in the unfertilised treatments in autumn 2012 (1.5

PRESS and Environment xxx (2014) xxx–xxx 7

years after the biochar application) provide partial evidence in sup-port of this mechanism in the present study. Nevertheless, neitherthe biochar-induced initial N immobilisation nor the increase insoil NO3

−-N contents in the second year were strong enough toalter the NU or grain yield. This is in agreement with a four-yearfield study on a fertile silt loam in central New York State, wherebiochar did not significantly affect the NU or grain yields of maize,but reduced N leaching by tripling the microbial biomass (Güerenaet al., 2012).

As found in the present study, the field-scale application of10–25 t ha−1 of biochar of woody raw materials has previously beenreported to increase the K content of non-weathered temperatesoils (Jones et al., 2012; Liu et al., 2012), whereas no significanteffect has been reported for easily soluble P, Ca, Mg and S 1–3 yearsafter application. The increased K content of the soil is attributableto the K in the biochar (Major et al., 2010; Quilliam et al., 2012;Xu et al., 2013; Tammeorg et al., 2014a), and there is yet no evi-dence of any longer-lasting differences in soil nutrient dynamicsin temperate and boreal fields (for instance, via increased cationexchange capacity of the soil). While the lack of differences in theeasily soluble P, Mg and S contents of the soil may be attributed tolow proportion of these nutrients in biochar, the contents of K andCa were both rather high. Even though the original soil was defi-cient in both of these elements, the soil content of easily solubleCa was about 20 times that of K, suggesting that the high levels ofbiochar-K may have displaced the Ca from soil. Similarly, Jones et al.(2012) reported 90% loss of the initial exchangeable K compared toonly 30% loss in exchangeable Ca of the biochar particles recoveredfrom a fertile sandy clay loam in Wales after a 3-year experiment.

Since B30 was associated with significant increases in WRC ofthe topsoil at 6 kPa and in AWC over control values in 2011, but B10was not, the effects of biochar on soil water properties appeared tobe dose-dependent. This is consistent with previous reports, andsuggests that biochar affected soil macropores more than micro-pores (Eastman, 2011; Liu et al., 2012; Abel et al., 2013). Similarly,in previous field-scale WRC measurements from undisturbed soilsamples, biochar application rates of 5–20 t ha−1 have been insuf-ficient for improving the WRC of a silt loam (Eastman, 2011),clayey Oxisol (Major et al., 2012), or sandy clay loam (Tammeorget al., 2014a), whereas at 25 t ha−1, biochar resulted in a significantincrease of WRC at 33 kPa and in AWC (difference between watercontent at 33 kPa and 1500 kPa; Eastman, 2011).

Further, the relatively high original SOM content of the soil(63.4 g kg−1) may explain the lack of effects of biochar applicationon the soil moisture content in any year, as the effects of biocharon soil water retention are less pronounced when SOM content isalready high (Abel et al., 2013). Abel et al. (2013) reported enhancedwater retention and AWC of laboratory-packed soil columns withlow SOM content soils (1–15 g kg−1), but no significant effect wasfound with biochar application to a soil with a high SOM content(91 g kg−1). Likewise, the combined application of 10 or 20 t biocharha−1 mixed with compost to a low SOM-content (16 g kg−1) loamysand increased the soil WRC significantly more than the applica-tion of compost alone (Liu et al., 2012), but the structure of thatexperiment did not allow separation of the interactive effects ofbiochar and compost from the main effect of biochar (Liu et al.,2012). The lack of biochar effects on soil physical properties in the2.5–7.5 cm layer in 2012, other than decreased bulk density andincreased porosity over the control, may be attributed to down-wards movement of biochar through tillage and root growth.

4.2. Growth, N uptake and yield of spring wheat

ort-term effects of biochar on soil properties and wheaton a boreal loamy sand. Agric. Ecosyst. Environ. (2014),

Unlike the fertilisers, biochar had no significant effects oncanopy growth dynamics, NU or the yield components of springwheat. This is in contrast with previous findings on Australian

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andy clay loams where wood biochar application at rates as lows 3 and 6 t ha−1 reduced drought stress and supported yield for-ation of wheat, an effect partly attributed to increased water

vailability through enhanced mycorrhizal colonisation (Blackwellt al., 2010; Solaiman et al., 2010). As the colonisation rate of AMas been reported to be inversely affected by soil P content (Jensennd Jakobsen, 1980; Ryan and Angus, 2003), any increase in AMolonisation would be smaller in our P-rich soil. In our previouseld study on a nearby boreal sandy clay loam, the use of 10 t ha−1

f biochar significantly increased the plant grain number of wheatn the dry year, 2011 (possibly due to relieved drought stress;ammeorg et al., 2014a). The high temperature and low precip-tation during flowering in 2011 decreased the moisture contentf the topsoil close to PWP and the plants in this study were pre-umably subjected to similar stress. The effect of biochar on soiloisture content was probably not sufficient to alter the yield for-ation, especially when the maximum biochar rate of applicationas equivalent to only 0.4% by weight of the soil to the 70 cm depth

rom which 95% of the water is typically extracted by wheat (Entzt al., 1992).

Similarly, since no rates of application had any effect on wheatrain yields in any fertiliser treatments, the biochar-induced pos-tive trends in SOC and available water contents were apparentlylso insufficient to affect yield in this soil with its rather high SOMontent. Furthermore, since the soil was nearly neutral (pH 6.4), thesual effect of biochar on reducing acidity (Major et al., 2010; Vanwieten et al., 2010; Vaccari et al., 2011) was not relevant. Althoughhe soil was initially considered deficient in several of the nutrientspplied with biochar (notably Ca, K, Mg and S), the lack of its effectn crop yields was consistent with previous 1–4 year studies on fer-ile soils in boreal (Karhu et al., 2011; Tammeorg et al., 2014a) andemperate climates (Güerena et al., 2012; Jones et al., 2012). Thisuggests that the biochar at the present application rates, irrespec-ive of its positive effects on K and NO3

−-N, did not sufficiently alterhe critical yield-limiting nutrient deficiencies. Further, the positiveorrelation between the grain yields and the content of soluble N,

and S in the soil suggests that these elements may have been theost limiting for the grain yields in this soil during these years.s the effect of biochar on crop yields has been shown to increasever time in previous studies (Steiner et al., 2007; Major et al., 2010;accari et al., 2011), there is a need for longer-term monitoring of

he nutrient release from biochar and consequent effects on plantrowth in the experiment.

.3. Organic vs. inorganic fertiliser

The low NU, SPAD and LAI values as well as grain yields androtein content in the MBM treatment suggest that this fertiliserade significantly less N available to plants than the inorganic fer-

iliser. This is in contrast with previous studies with spring wheatn Norwegian silt loam (Jeng et al., 2004) and with barley and oatsn Finnish silty clay loam (Chen et al., 2011), and may be attributedo initially lower available N content of either the soil or the MBM.n the other hand, the constantly increased easily soluble P contentnd initially incremented Ca and S contents of soil over the inor-anic fertiliser treatments could be attributed to somewhat greaterineralisation of MBM-P than the suggested 18% (Ylivainio and

urtola, 2009), as well as to the greater amounts of Ca and S in theBM than in the inorganic fertiliser.Apart from the signs of initially increased N immobilisation, the

ffects of biochar on the soil chemical properties and wheat yieldormation did not show significant differences between MBM and

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norganic fertiliser treatments. This suggests that if biochar is useds a C sequestration tool in combination with either inorganic orow C:N ratio organic fertilisers like MBM, no negative effects on soilhemical properties or in yield formation of spring wheat should be

PRESS and Environment xxx (2014) xxx–xxx

expected at rates up to 30 t ha−1. Further research is needed to seewhether interactive mechanisms not studied in this study, such asa shift in soil microbial community towards bacterial dominanceover fungi as reported separately for both MBM (Mondini et al.,2008) and biochar (Jones et al., 2012), will contribute to agronomicbenefits in boreal climate over a longer time span.

5. Conclusion

This 2-year field study under boreal conditions showed that theapplication of spruce chip biochar to a nutrient-deficient loamysand, either alone or combined with inorganic fertiliser or MBM,had no negative impacts on soil physicochemical properties or theyield and quality of spring wheat, apart from short-term N immo-bilisation in the first year. On the contrary, in the second year thehighest biochar application rate (30 t biochar ha−1) decreased thebulk density and increased both the porosity and the NO3

−-N con-tent of the soil over the control. These effects, together with durablyincreased soil K and initially increased SOC content, WRC and AWC,were, irrespective of the type of the fertiliser, insufficient to influ-ence the yield formation and the grain yield. This was possiblydue to low availability of biochar nutrients and the relatively highunderlying SOM content of the soil reducing the effects of biocharon soil water retention. We also found that only 18% of the C addedin the biochar could be detected in the topsoil two years after itsapplication. The reasons behind this, as well as longer term changesin nutrient availability and plant growth, remain to be studied inlonger term experiments.

Acknowledgements

We thank Sampo Tukiainen (Preseco Oy) for providing thebiochar, Miia Collander and Johanna Muurinen for the technicalassistance with biochar and soil analysis, Mikko Hakojärvi for hishelp with the TDR measurements and Markku Tykkyläinen for histechnical assistance in the field. This study was funded by Jennyand Antti Wihuri Foundation and the Ministry of Agriculture andForestry of Finland.

Appendix A. Supplementary data

Supplementary data associated with this article can be found, inthe online version, at http://dx.doi.org/10.1016/j.agee.2014.01.007.

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