Plant Soil DOI 10.

1007/s11104-013-1745-6

REGULAR ARTICLE

Degradation kinetics of biochar from pyrolysis and hydrothermal carbonization in temperate soils
Mo Bai & Burkhard Wilske & Franz Buegger & Jrgen Esperschtz & Claudia Irene Kammann & Christian Eckhardt & Martin Koestler & Philipp Kraft & Martin Bach & Hans-Georg Frede & Lutz Breuer

Received: 5 October 2012 / Accepted: 26 April 2013 # Springer Science+Business Media Dordrecht 2013

Abstract Background and Aims Estimates of biochar residence times in soils range over three orders of magnitude. We present the first direct comparison between the biodegradation of a char from hydrothermal carbonization (htcBC) and pyrolysis (pyrBC) with high temporal resolution. Methods Mineralization of the biochars and their shared Miscanthus feedstock in three soils was determined directly by the 13CO2 efflux using a novel method incorporating wavelength scanned cavity ring-down spectroscopy. Biochar half-life (t1/2) was estimated with three empirical models. Results (1) The htcBC was readily biodegradable, whereas pyrBC was more recalcitrant. (2) Cumulative

degradation of both biochars increased with soil organic carbon and nitrogen content. (3) The corrected Akaike information criterion (AICC) showed an overall preference for the double exponential model (DEM) reflecting a labile and a recalcitrant C-pool, over the first-order degradation model (FODM) and a logarithmic model. (4) The DEM resulted in t1/2 ranging from 19.744.5, 0.72.1 and 0.81.3 years for pyrBC, htcBC and feedstock, respectively. Conclusion The degradation was rather similar between feedstock and htcBC but one order of magnitude slower for pyrBC. The AICC preferred FODM in two cases, where the DEM parameters indicated no distinction between a labile and recalcitrant carbon pool. Keywords Char . HTC . Soil amendment . Recalcitrant carbon . Biodegradation . 13CO2 efflux

Responsible Editor: Eric Paterson. M. Bai : B. Wilske (*) : M. Koestler : P. Kraft : M. Bach : H.<G. Frede : L. Breuer Institute for Landscape Ecology and Resources Management (ILR), Research Centre for BioSystems, Land Use and Nutrition (IFZ), Justus Liebig University, Gieen, Germany e-mail: brkwils@yahoo.com F. Buegger : J. Esperschtz Institute of Soil Ecology, German Research Center for Environmental Health, Helmholtz Zentrum Mnchen, Neuherberg, Germany C. I. Kammann : C. Eckhardt Department of Plant Ecology, Research Centre for BioSystems, Land Use and Nutrition (IFZ), Justus Liebig University, Gieen, Germany

Introduction Biochar has been reinvented as a multi-tool raising expectations to enhance bioenergy production, soil water availability, crop growth, and the carbon sequestration of soils (Glaser et al. 2000; Lehmann 2007; Gaunt and Lehmann 2008; Atkinson et al. 2010; Zimmerman 2010; Hammond et al. 2011; Matovic 2011). Most of the recent studies were fairly positive suggesting that biochar amendments can indeed (a) improve the water availability in soils (Karhu et al. 2011), (b) increase both the water use efficiency and

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nitrogen use efficiency and thereby the growth of plants (Kammann et al. 2011a; Prendergast-Miller et al. 2011; Vaccari et al. 2011), (c) reduce emissions of major greenhouse gases (e.g., N2O, CH4) from agricultural land (Karhu et al. 2011; Kammann et al. 2011b; Knoblauch et al. 2011), and (d) amplify the pool of recalcitrant carbon (C) in soils to enhance their C sequestration (Vaccari et al. 2011; Knoblauch et al. 2011). Other effects of biochar include the reduction of herbicide efficiency (Nag et al. 2011) and the increased retention of heavy metals, respectively (Cao et al. 2011; Uchimiya et al. 2011; Buss et al. 2011). Few observations point to possible caveats in our understanding of biochar functions in the terrestrial environment, e.g., one study suggested increased humus loss in the presence of char from wildfire (Wardle et al. 2008). Biochar includes a wide spectrum of materials with a large variability of individual properties (e.g., Lehmann 2007; Libra et al. 2011; Liu et al. 2010). Some properties, or the modifications of the same, are dependent on the conversion process from biomass to char (Schimmelpfennig and Glaser 2011). Many studies have considered biochar from pyrolysis (hereinafter pyrBC), which represents the traditional thermal conversion without oxygen. Hydrothermal carbonization (HTC), also termed wet pyrolysis (Libra et al. 2011), is a relatively new process providing biochar (hereinafter htcBC) from a slurry of biomass and water subjected to autogenous pressure (e.g., 1 MPa at about 200 C). With respect to the variety of available biomass, HTC complements the process of pyrolysis, because htcBC can be produced from biomass involving much higher water contents. Similar to pyrBC, htcBC has been considered a potentially useful soil amendment (Libra et al. 2011; Kammann et al. 2011b), and positive effects on the root colonization of fungal symbionts were reported (Rillig et al. 2010). However, residence times of pyrBC and htcBC must be studied in detail to pursue long-term C sequestration (Fuertes et al. 2010). This expands to a greater task, as the property of biochar stability (or residence time in soil) does not only depend on the feedstock and production process, but also on the soil conditions (Zimmerman et al. 2011). While a large variety of biochar products is prepared to enter the market (Meyer et al. 2011), the estimates of biochar residence times in soils stretch over three orders of magnitude (i.e., 41,000 years) (Lehmann et al. 2009; Kuzyakov et al. 2000; Steinbeiss et al. 2009; Cheng et al. 2008). Hence, efficient methods are required to

investigate the stability and/or the C sequestration potential of many different biochars in a variety of soils. The objectives of the present study were to (1) investigate the biodegradation of one pyrBC, one htcBC, and their shared feedstock (Miscanthus) in relation to the incubation in three common agricultural soils, and (2) estimate the half-life of biochar based on existing models that were previously applied to describe its mineralization. To our knowledge, this is the first direct comparison of the biodegradation of these two profoundly different biochars. At the same time, we present a novel and efficient method to measure biochar degradation. Biodegradation of the pyrBC and the htcBC was based on the 13CO2 efflux measurements from biochar-soil incubations using wavelength-scanned cavity ring-down spectroscopy (WS-CRDS). Both 13C- and 14C-labeling can be employed to measure CO2 evolution from biodegradation of recalcitrant carbon compounds such as black carbon or biochar (Kuzyakov et al. 2009; Jones et al. 2011). However, the 13C-labeling has the advantage of not being subjected to special laboratory standards and environmental concerns. Coupling the WS-CRDS technique to auto-sampling (Bai et al. 2011), we were able to directly measure 13CO2 emissions from a series of sample incubations. The novel method increases the frequency and flexibility in accessing degradation rates with high accuracy, and it reduces simultaneously the systematic or shifting errors owing to sample handling. A temporal resolution of weekly measurements provided the basis to examine the degradation kinetics and to calculate biochar half-lives (t1/2) as a benchmark for the related carbon sequestration potential (Zimmerman 2010; Gaunt and Lehmann 2008; Qayyum et al. 2011). Three models were used to estimate half-life: (1) the first-order decay rate model (FODM, Bolan et al. 2012), (2) the double exponential model (DEM, Lehmann et al. 2009; Zimmerman et al. 2011), and (3) a logarithmic model (LOGM, Zimmerman 2010). Anderson et al. (2011) suggested that the degradation of more recalcitrant carbon pools requires sufficient diversity within the soil microbial population. Although biochar can increase soil microbial activity and affect soil enzyme activities (Bailey et al. 2011; Wardle et al. 2008), the microbial activity prior to any treatment depends greatly on native soil organic carbon (SOC) (Pascual et al. 1997; Insam and Domsch 1988). As litter-derived inputs to the three soils were equivalent over many years, we suggest that main

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differences between soils were in SOC amount, rather than SOC quality. Therefore, the degradation kinetics of the chars were expected to reflect biochar soil interaction, with SOC content and/or microbial activity being the key players. Thus, we set out from the hypothesis that for each of the biochars, the rate of degradation was mainly proportional to the different SOC contents of the three soils tested.

Biochar degradation Degradation is the successive breakdown and mass loss of a substance and biodegradation implies the same is caused by biological means (organisms, enzymes, etc.). Biodegradation of a substance in a matrix such as soil will thus depend on the soil biological activity, particularly if the substance can be presumed being not readily biodegradable. As a first approximation, biological activity in turn will depend on substrate availability, i.e. the amount of SOC (Insam and Domsch 1988). Hence, native SOC was used as a basis to adjust the amount of biochar application to the soils. In each treatment biochar equal to 50 % of the native SOC was mixed with 300 g soil (corresponding to 3.4, 9.7 and 5.0 g C kg1 added to soils in LUFA 1, 2 and 3, respectively). The biochar added corresponded to 19 to 66 t ha1 depending on the different soils. These amounts comply with the German by-law for the application of biodegradable waste to agricultural land (Bioabfallverordnung 2012), which provides ranges equaling 2075 % of the native SOC. Biochar incubation The two biochars and the feedstock were incubated with the three soils over a period of 200 days. Samples were transferred to 1-L vessels and incubated constantly at 25 C and 40 % soil water holding capacity (WHC). The water content was adjusted weekly. Between measurements, the samples were kept in a dark climate chamber at 70 % relative air humidity to avoid photosynthetic activity and drying of soils. Samples of pure soil were used as references, i.e. the 13CO2 efflux of the reference was subtracted from the treatment to obtain the 13CO2 net efflux. Each treatment in each soil ( n = 3 3), and the reference samples for each soil (n =3), were tested in four replicates (total n =48). Measurement of biochar degradation Biochar degradation was measured based on the 13 CO2 efflux from the samples each day from day 1 to day 7, and every week thereinafter up to 200 days. The study was conducted with a novel automated system that couples a batch of samples via two rows of microprocessor-controlled valves to a WS-CRDS

Materials and methods Biochars and soils Pyrolysis biochar and char from hydrothermal carbonization were produced from straw feedstock of Miscanthus x giganteus at highest temperature treatments of about 57525 C and 200 C, respectively (Table 1). Total process time was 30 min for the pyrBC, whereas process time and pressure were 120 min and 1.6 MPa for the htcBC. The pH and ash content of both feedstock and biochars were measured according to van Reeuwijk (2002) and based on the loss on ignition method (Heiri et al. 2001), respectively. Processing increased the pH from feedstock (6.8) to htcBC (10.1) but lowered it to pyrBC (5.1). The ash content increased from feedstock (2.5 %) to htcBC (2.7 %) and further to pyrBC (29.8 %). Utilization of the C4 species Miscanthus significantly enriched the 13C content in the biochars as compared to the soils used for incubation (Table 1). The 13C and organic carbon contents of the biochars, the feedstock, and the soils were determined using an IR-MS (delta V Advantage, Thermo Finnigan, Bremen, Germany) coupled with an elemental analyzer (Euro EA, Eurovector, Milano, Italy) at the Helmholtz Centre Munich. Each treatment, the two biochars and the feedstock, was crushed to a particle size 1.5 cm. Three soils were obtained from the state research institute for agriculture (Landwirtschaftliche Untersuchungsund Forschungs-Anstalt LUFA Speyer; www.lufaspeyer.de). The soils from LUFA 1 to LUFA 3 represent a gradient from sand to loam texture. In contrast, both soil organic carbon and nitrogen (N) contents increase from LUFA 1 to LUFA 3 to LUFA 2 (Table 1). The soils were air dried and passed through a 2-mm sieve.

Plant Soil Table 1 Properties of the LUFA soils 13, Miscanthus feedstock, and the biochars from pyrolysis (pyrBC) and hydrothermal carbonization (htcBC) Property Soil LUFA 1 Inceptisol Soil texturea Corg (%) Total N (%) C/N ratio 13C Sand (%)a Silt (%)a Clay (%) pHb,c HTTd (C)a Process. (min)
f a a

Feedstock LUFA 2 Mollisol Loamy sand 1.93 0.17 11 27.8 81.3 12.1 6.6 5.1 <2 45.2 1247 LUFA 3 Inceptisol-Aquept Sandy loam 0.99 0.08 12 25.2 62.5 28.7 8.9 5.5 <2 35.6 1291 48.95 0.17 295 11.9 6.8 2.5 <15 Miscanthus

Biochar htcBC pyrBC

Sand 0.68 0.04 17 27.6 87.6 9.3 3.0 7.1 <2 31.4 1455

52.30 0.31 169 12.4 10.1 200 120 2.7 <15

76.10 0.63 121 14.2 5.1 575e 30 29.8 <15

Ash content (%) Particle size (mm) WHCg (%)a Bulk density (g/L)a
a b c d e f g

Parameter values as per the supplier Soil pH (0.01 M CaCl2) pH of biochar and feedstock in H2O HTT: Highest temperature treatment +/25 C Processing time Water holding capacity

CO2 analyzer (G1101-i, Picarro, Sunnyvale, CA, USA). Details of the method and the system have been described elsewhere (Bai et al. 2011). Briefly, the WSCRDS technique provides direct quantification of the 13 C stable isotope in the sample gas, i.e. the mixing ratio of 13CO2 (mol mol1). The recently developed method is based on the open dynamic chamber mode which has proven to provide reliable results to determine CO2 efflux from soils (Baldocchi and Meyer 1991; Pumpanen et al. 2004). Samples are consecutively connected through to the WS-CRDS, while the other samples are flushed with ambient air by an external pump at the same rate. When a sample is connected to the analyzer, carry-over effects are flushed out during the first 180 s, and the 13CO2 efflux is averaged over six records during the following 60 s. Subsequently, the 13CO2 concentration of the ambient air is measured for 180 s. Calibration of the measurements was checked regularly using at least two 13CO2

13

isotope standards enclosing the target mixing ratios (e.g., 20 V-PDB in CO 2 totals of 200 mol mol1 and 1,000 mol mol1; DEUSTE Steininger GmbH, Mhlhausen, Germany). Calculation and statistics The 13CO2 efflux rate of both the treatment (soil+ biochar) and the reference sample (soil only) was calculated as the difference between the 13CO2 concentration in ambient air before and after passing the chamber. The 13CO2 efflux rate F (g d1) is F U 106 Cout Cin b 60 24 MV 1

where is U the gas flow rate with 22 mL min1, Cout (mol mol1) the 13CO2 concentration in the chamber, Cin (mol mol1) the 13CO2 concentration in ambient air (mol mol1), b the molar mass of 13C (g mol1),

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and MV the temperature- and pressure adjusted molar volume (m3 mol1). The initial weight of soil was similar in sample and reference, and hence, the net 13 C efflux rate Fnet (g d1) due to biochar degradation is Fnet Ftreatment Fsoil
13

where Ftreatment and Fsoil are C efflux rates (g d1) from treatment and reference soil, respectively. Fnet (g d1) was referenced to the initial 13C application (g) resulting in Fdeg (g g1 d1) by  c  1 Fdeg Fnet a 100     e 1 1 d 1 3 1000 where is a the biochar initial weight (g), c the biochar organic carbon content (%), d the 13C/12C-ratio of the international standard VPDB (i.e. 0.0111802) (Werner and Brand 2001), and e the biochar 13C. For further analysis, the mean degradation curve from the efflux of degraded 13C of four replicates per day was used to fit different equations to describe C degradation. The cumulative 13 C mineralization 13 Clost: 
13

biodegradation of poultry manure compost and poultry manure biochar, respectively. (2) A double exponential model (DEM; Eq. 7) was used to estimate the mineralization of both an htcBC (Qayyum et al. 2011) and pyrBCs (Lehmann et al. 2009; Zimmerman et al. 2011) assuming there are two carbon pools involved: One rapidly degrading C-pool and one slowly degrading or recalcitrant C-pool. (3) Zimmerman (2010) proposed a logarithmic model (LOGM, Eq. 9) to predict the long-term biodegradation of pyrBC. The FOD model is:
13

Crem;FODM 13 C0 ek t
13 13

where Crem,FODM is the C weight remaining from the initial weight after a certain number of incubation days and the value k is the rate constant of decrease (d1). The DEM considers two carbon pools 13C1 and 13 C2 with different turnover rates k1 and k2, respectively:
13

Crem;DEM 13 C1 ek1 t 13 C2 ek2 t

Clost 13 Ct1

Fdeg;t Fdeg;t1 2
1 13

 t t 1 4

If k is known, the half-life t1/2 can be calculated with Eq. (8) for the FODM and DEM, respectively. The value k2 can be used for the half-life calculation based on DEM, if C2 >> C1 and k2 << k1, (Qayyum et al. 2011). The biochar half-life (yr) is: t1=2 ln2 k 365 8

where Clost (g g applied C) is the cumulative 13 C mineralization after t (days), 13Ct1 is the cumulative 13C mineralization of the previous day, Fdeg,t is the degradation 13C in t days, and Fdeg,t1 is the degradation of the previous day. Note that the mineralization between two sampling is assumed as linear. For the mathematical description of biochar degradation the carbon remaining (13Crem, calculated to g g1) from the initial carbon applied (13C0) at the start of experiment is required. The remaining 13Crem in initial weight is:
13

The two following equations (Eqs. 9 and 10) were used to calculate the cumulative C mineralization 13 Clost after any given period time t, and the half-life based on the LOGM. The term for 13C mineralization is:  13  C 0 eb 13 Clost 9 t m 1 m1 The term for half-life estimation based on LOGM is:  t1=2 m1 2 eb   
1 m1

13

Crem 13 C0 13 Clost

From this, the half-life of biochar in years t1/2 (yr) can be computed. We compared three models suggested by previous studies: (1) Bolan et al. (2012) used the first-order decay rate equation (FODM; Eq. 6) to model the rapid and slower

10

where m and b are calculated via regression analysis with ln(Fdeg/365)=m ln(yr)+b, and Fdeg is 13 C degradation rate (g g1 13C d1). Note that

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the degradation rate has to be converted to time steps as fraction of year (yr). The degradation rates of biochar and feedstock were analyzed using the repeated measurement analysis of variance (RM-ANOVA). The soil type, treatment and time were used as fixed factors and their interaction. The normal distribution for all variables was tested using the Shapiro-Wilk test. Two factorial ANOVA was used to distinguish between htcBC, pyrBC and feedstock. We analyzed results of the same soil or tested between three soils containing the same application at 200 days of the incubation (p <0.05). The two factors were soil type and treatment, i.e. the addition of different biochars or feedstock. Non-linear regression and linear regression analyses were used to calculate the parameter k for both FODM and DEM, and m and b for LOGM, respectively. Statistics were computed using SPSS (IBM SPSS Version 20, NY, USA). Root mean square deviation (RMSD) was calculated to characterize the goodness of fit between measured and modeled data. The Akaike information criterion (AIC) was used to indicate the model performance, i.e., the goodness of model fit in relation to the number of estimated model parameters, of the three models for each treatment (Akaike 1981). AIC values provide a mean for model selection by weighing the goodness of fit versus the number of parameters included in a model, thereby penalizing overfitting. We used the corrected AIC (AICC), which corrects for small n (Burnham and Anderson 2004). AIC can range from positive to negative values, whereby the lowest value indicates the best approximating model.

Results The degradation of biochars The degradation was significantly different between feedstock and pyrBC, and between htcBC and pyrBC, respectively after 200 incubation days (p <0.0001, Fig. 1). Until at least day 100, degradation rates of Miscanthus feedstock and htcBC were mostly similar in the LUFA 1 soil (p =0.84) and from day 7 to day 100 in the LUFA 2 soil (p =0.18) (Fig. 1a, b). Only in the LUFA 3 soil, degradation was larger for the feedstock than the htcBC (p <0.0001). After 100 days, the slope of htcBC degradation curves flattened, i.e. the degradation rates decreased, as compared to the curves
Fig. 1 Remaining 13C in initial weight of the applications (13Crem) of feedstock (Miscanthus), htcBC and pyrBC in three LUFA soils

of the feedstock (Fig. 1b). Among the htcBC curves, the htcBC degradation in LUFA 1 showed another flattening of the slope after about day 150. As a result, the mean 200 day degradation rates for htcBC were 269.7 mg g1 (27 %), 304.9 mg g1 (30 %), and 299.4 mg g1 (30 %) in LUFA 1, LUFA 2, and

Plant Soil Table 2 Cumulative 200 day biodegradation of BC and feedstock (mg13C g1 applied Soil Miscanthus Biochar htcBC LUFA 1 LUFA 2 LUFA 3 323.7 346.7 367.3 7.7 a 18.2 19.6 b
A 13

C). Values are mean SD (n =4, p <0.05)

pyrBC 10.0 a 11.1 13.9 b

B

269.7 304.9 299.4

5.6 25.8 11.4

7.4 a 4.3 3.4 c

ab A A

b B B

b C C

Capital letters indicate significant differences in results between BC and feedstock relative to the same soil; small letters indicate significant differences between similar applications (BC type or feedstock) in different soils, respectively

LUFA 3, respectively (Table 2). The mean 200 day degradation rates for the feedstock were slightly higher with 32 %, 35 %, and 37 % in LUFA 1, 2, and 3, respectively. The degradation of pyrBC within the three soils showed diverging curves (Fig. 1c). After 3 weeks of incubation, pyrBC degradation was clearly larger in LUFA 2 than in the two other soils. Virtual increases in initial weight of pyrBC in LUFA 1 and 3 (Fig. 1c) were a result of repeated negative efflux rates, i.e. the efflux from the treatment sample was smaller than from the reference sample. The mean 200 day degradation rate of pyrBC was 25.8 mg g1 (<3 %) and 11.4 mg g1 (1 %) in LUFA 2 and LUFA 3, respectively (Table 2). The 200 day mean degradation of pyrBC in LUFA 1 resulted in a negative value (5.6 mg g1), which means that less 13C was degraded from the pyrBC samples than from the reference (soil only). However, the mean was not significantly different from zero. The two factorial analysis confirmed that the 200 day degradation rates were different between both the biochar types and the biochars in different soils, whereas there were no significant differences from the interaction of soils and biochar types (p = 0.059, Table 2). In each of the three soils, the 200 day degradation of htcBC was significantly smaller, although only by 1218 %, than those of the feedstock. In contrast, the pyrBC degradation rates were an order of magnitude lower as compared to those of htcBC. The 200 day degradation of Miscanthus feedstock appeared to increase along the texture gradient from sand to sandy loam (LUFA 1 to 2 to 3), with the LUFA 2 result being not significantly different from either of the two others (Table 2). The degradation of both biochars increased with increasing native SOC and N

content from LUFA 1 to 3 to 2 (Table 1). This result was significant throughout for pyrBC (Table 2), whereas for htcBC, it was only significant for the step from LUFA 1 to LUFA 3. This first increase in both SOC and N was relatively smaller than the second from LUFA 3 to 2, but it marked the larger decrease in C/N ratio (Table 1). The fit of model data Employment of the three kinetic models FODM, DEM and LOGM resulted in a superior curve fitting, i.e., the lowest RMSDs, for the DEM across the three soil and two BC types (Fig. 2af). Furthermore, RMSDs were lower for the FODM than the LOGM in all but one case. For the htcBC in LUFA 1 (sand), FODM and LOGM overestimated the degradation by about 0.05 g g 1 13 C application (RMSD=0.022) and 0.1 g g1 13C application (RSMD = 0.127) as compared to measured data, respectively (Fig. 2a). Also for the degradation of htcBC in LUFA 2 (loamy sand) and LUFA 3 (sandy loam), the DEM model matched the measured data best; although differences in RMSD between the three models were less pronounced as compared to LUFA 1 (Fig. 2b, c). The absolute differences in RMSD were generally smaller in case of the pyrBC applications (Fig. 2df), and FODM and LOGM performed almost equal as DEM in case of LUFA 2 and 3. However, the DEM model showed a substantially lower RMSD with 0.001 for LUFA 1, where especially LOGM overestimated the degradation of biochar. Only from the charts but not the RMSD it became obvious that some differences between measured and modeled increased towards the end of the observation period (see e.g., Fig. 2b, DEM).

Plant Soil Fig. 2 Comparison between the remaining 13C in initial weight (13Crem) of htcBC and pyrBC in three LUFA soils based on the average measured degradation curve, the first order degradation model (FODM), the double exponential model (DEM) and the logarithmic model (LOGM). RMSD is the root mean square deviation between measured and modeled data

The half-life of biochar Calculated half-lives of htcBC in three soils, based on the three models FODM, DEM and LOGM, ranged between 0.7 and 2.1 years (Table 3). They were only slightly higher than the half-lives of pure Miscanthus feedstock, ranging between 0.7 to 1.3 years. In most cases, LOGM estimated a 4 month longer half-life than the FODM model for htcBC. The DEM, which was fitting best to the existing data of htcBC degradation, suggested halflives of 0.7, 1.1, and 2.1 years for htcBC in LUFA 1, LUFA 2, and LUFA 3, respectively (Table 3). In sand, loamy sand, and sandy loam the t1/2 results compared between the models as LOGM > FODM > DEM, LOGM > DEM > FODM, and DEM > LOGM > FODM, respectively.

Model predictions for the half-life of the pyrBC ranged between 11.1 and 110.7 years (Table 3, LOGM). Overall, the results for t1/2 of pyrBC increased from LUFA 2 (loamy sand) to LUFA 3 (sandy loam) to LUFA 1 (sand), along with the decrease in SOC and N content. Results of pyrBC t1/2 using all three models for the different soils varied in rough comparison between 11 and 15 (LUFA 2), 1945 (LUFA 3) and 20111 years (LUFA 1). In one and two cases, the k value of FODM and the k2 values of DEM were not significantly different from zero, respectively. Overall, the three models did not reveal a similar and thus clear pattern with regard to estimated half-lives of pyrBC in the different soils. However, we found a clear tendency for substantially longer halflives of pyrBC in relation to htcBC and Miscanthus feedstock in the order of a magnitude and even more.

Plant Soil Table 3 Model parameters and half-lives of biochar and Miscanthus feedstock in three soils. Half-life (t1/2, yr) is estimated by three models (first-order degradation FODM, double Soil Application FODM k AICc t1/2 DEM C1 k1 C2 k2 AICc t1/2 412 376 exponential DEM, and logarithmic model LOGM). Model performance is assessed by the corrected Akaike information criterion (AICc) LOGM m b AICc t1/2 1.0 1.4

LUFA 1 Miscanthus 2.16E-03 307 htcBC pyrBC 1.95E-03 249

0.9 9.19E-05 0.03a 1.0 7.66E-03 0.02a

0.99 2,30E-03 0.99 2.60E-03

0.8 0.35 1.13 255 0.7 0.73 2.10 129

8.41E- 413 b 0.01 06a LUFA 2 Miscanthus 2.19E-03 311 0.9 htcBC pyrBC htcBC pyrBC Note that C2 =1 C1
a b c d

6.64E-03a 0.99 9.64E-05a d 0.10 1.26E-03 0.02 0.01 d 1.00 2.19E-03 0.96 1.74E-03 0.84 1.44E-03 0.80 8.91E-04 1.00 4.27E-05

467 19.7 0.29 4.14 300 110.7c 306 346 408 371 0.9 0.29 0.96 296 1.1 0.42 1.39 277 0.10 3.38 411 1.3 0.44 1.17 223 2.1 0.44 1.41 251 0.36 3.08 382 0.9 1.3 11.1c 0.8 1.3 18.7c

2.03E-03 278

0.9 0.04 0.7 0.16 0.9 0.20

1.31E-04 460 14.5 0.17 2.03E-03 275

0.83 8.30E-05a 468 b

LUFA 3 Miscanthus 2.56E-03 258

4.27E-05 418 44.5

413 44.5

Not significantly different from 0 Not calculated because k <0 or k2 <0 Not reliable based on Zimmermann (2010) since linear regression yields R2 <0.4 Because k1 identical to k2

The AICC values assessed better model performances for the DEM than the FODM, and further, better performances for the FODM than the LOGM (Table 3). DEM and FODM showed the best performance in seven of nine and two of nine cases, respectively. In those two cases, where the DEM performance was only second best, the k-values for the labile and recalcitrant pool were identical. While the LOGM performance was always assessed the least best among the models, the difference in AICC to the next better one ranged between 1 and 120 points.

Discussion Biochar properties and degradation The 200 day degradation of both feedstock and htcBC were one order of magnitude larger than the degradation of pyrBC in all of the soils tested. In contrast, the degradation was not significantly different between feedstock and htcBC in two of three soils. Among the property data available for the feedstock and the biochars (Table 1), it was the ash content that correlated with the large difference in degradation between

pyrBC and the other two treatments. Also Chen et al. (2008) found that ash content increases linearly with pyrolysis temperature for pyrBC derived from pine needle litter. Spokas (2010) suggested H/C and O/C ratios as stability indicators, both of which decreased linearly with increasing ash content. Previous studies have also shown that both H/C and O/C ratios were much lower in pyrBC than in htcBC (Fuertes et al. 2010; Schimmelpfennig and Glaser 2011). Further parameters mainly showed a gradual increase (Corg, total N) and decrease (C/N ratio) from feedstock via htcBC to pyrBC, and no clear relationship with biochar degradation. Overall, these relations suggest that the much slower degradation of pyrBC as compared to htcBC was related to a greater extent of carbonization (Liu et al. 2010) including higher ash content and lower H/C and O/C ratios. Biochar degradation in different soils A recent study proposed that environmental and biological controls rather than material properties predominate in controlling the stability of soil organic matter (Schmidt et al. 2011). The results of the present study showed differences between the 200 day

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biodegradation rates of the feedstock, the htcBC and the pyrBC in the different soils. Initially high rates of degradation were observed for both chars and were probably due the decomposition of remaining volatiles and soluble compounds (Luo et al. 2011; Smith et al. 2010). In spite of the differences in the magnitude of the 200 day degradation, the degradation of both htcBC and pyrBC increased with increasing native SOC and N content. The degradation curves of htcBC were very similar in their course (Fig. 1). Where they diverged, it was between the application in LUFA 1 and the other two soils. Both SOC and N content changed along the sequence LUFA 1 to 3 to 2. However, the largest difference between LUFA 1 and the other two soils was the wider C/N ratio. This relationship suggests htcBC degradation is likely to increase following N-fertilization. Degradation of pyrBC was very low (LUFA 2 and 3) or not detectable (LUFA 1), but significantly different between LUFA soils and also to htcBC (Table 2). Luo et al. (2011) studied the biodegradation of 700 C and 350 C pyrolysis biochar (pyrBC700, pyrBC350) produced from Miscanthus. They report a 87 day biodegradation of 0.14 % and 0.18 % of pyrBC700 and 0.61 % and 0.84 % of pyrBC350. At day 91, the pyrBC used in the present study (produced at a temperature of 575 C) showed a similar, low degradation of the same magnitude with 0.11 %, 1.24 % and 0.31 % in the LUFA soils 13, respectively. Incubation of pyrBC in LUFA 1 even lead to negative degradation rates, i.e. a higher 13CO2 emission of the control compared to soil incubated with pyrBC. Negative degradation rates can be explained as following: the application of 13C enriched C4-plant derived biochar (in our case Miscanthus biochar) usually results in increased 13CO2 emission rates relative to a control soil (in our case a soil that is characterized by plant residues stemming from C3-plants). Higher emission rates of the control compared to that of the biochar amended soil can be explained by a suppression of the microbial degradation of native SOC, e.g. through N immobilization (Zavalloni et al. 2011), adsorption of soil-borne CO2 or of otherwise easy degradable volatile organic compounds. Previous studies suggested that pyrBC produced at high temperatures can exert negative priming effects during early and later periods of incubation (Zimmerman et al. 2011; Luo et al. 2011; Cross and Sohi 2011), an effect that could explain our observations. A clear

separation of the different carbon pools (here native SOC and biochar) contributing to observed 13CO2 emissions is not possible following the analytical method by Bai et al. (2011), which we followed in this study. Applying well-known mixing model approaches using 13C signatures (Paterson et al. 2009) could resolve this limitation. But this is currently not feasible with the first generation WS-CRDS analyzer we used here due to the relative low precision of the 13C signal provided by the analyzer. In theory, the analyzer could provide higher accuracies, but for this longer incubation times of up to 1 h are needed, contradicting our aim of rapidly measuring a large number of samples (Bai et al. 2011). For further details see the recently published performance check of the Picarro G-1101-i analyzer by Vogel et al. (2013), the same instrument we used here. Nevertheless, all this is not limiting the value of our study as we do not focus on the investigation of priming effects of biochar application but rather on the relative degradation of different biochar types relative to their feedstock in different soils. Increasing biochar degradation from LUFA 1 to 3 to 2 did not correlate with the texture or the clay content of the soils. Similarly, Saggar et al. (1996) found that degradation of organic matter in soil was not correlated to the clay content but related to the surface area of clays. Hassink (1994) reported that the microbial biomass activity in clay was lower than in sandy and loamy soil. Baldock and Skjemstad (2000) linked the two prior findings by suggesting that the best condition for the decomposer consists of good water supply and sufficient oxygen supply through open pore space. Reviewing mechanisms for black carbon stabilization in soil, Knicker (2011) considered that association and/or microaggregate formation with soil minerals can provide another protection from degradation. Thus, although the clay contents were relatively low in all of the three LUFA soils, a retarding effect of biochar degradation through its interaction with clay minerals may be still possible over longer time periods. The rapid degradation of the tested htcBC suggests that it is not suitable for enhancing the long-term carbon sequestration of soils (Woolf et al. 2010). The htcBC might still be considered a valuable soil amendment with respect to its higher N-content as compared to the feedstock. However, a recent study pointed to the htcBC

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treatments potentially including the risk of increased N2O emissions (Kammann et al. 2011b), which lowers expectations htcBC might involve generally reduced emissions of potent greenhouse gases as compared to unprocessed biomass. Measuring biochar degradation A general standard method has yet to be defined for the analysis of biochar degradation in soil (Koide et al. 2011). The present study used a method, which facilitates characterization and comparison of the degradation of diverse biochar products under constant temperature and moisture conditions. It is well suited for standardized, early assessment of biochar degradation and to complement long-term experiments under field conditions (Bai et al. 2011). Soil moisture is a major regulator of seasonal microbial biomass in soil samples (Van Gestel et al. 1992), and proportions of biochar incubation in soil and the soil water content applied were similar to other studies (Luo et al. 2011; Peng et al. 2011). The temperature and constant soil moisture conditions of the present laboratory study were conducive to biodegradation, which suggests that degradation rates are overestimated when compared to the seasonally varying conditions in the field (Kuzyakov et al. 2009). While supporting the characterization of the individual biodegradability of different biochar products, the degradation rates reported in this study may not relate directly to those under field conditions. Field amendments of biochar are usually mixed with manure or the like to avoid nutrient capturing by fresh char (PrendergastMiller et al. 2011; Gaji and Koch 2012). Also annually renewed inputs of organic matter and nutrients from biomass have far-reaching effects on biochar degradation (Kammann et al. 2011b). There are also limits to the period, over which the results of a laboratory study can be useful to draw conclusions on biodegradation in-situ. For example, differences to field conditions may increase due to the depletion of microbial composition and competition during long-term incubation and without seasonal changes in soil temperature, and triggering cycles of dry-wet conditions (Potts et al. 2006). Overall, the multitude of drivers suggests that the average field conditions do not exist, and the extent to which laboratory results over- or underestimate half-lives of biochar in the field varies from site to site and or year to year.

Biodegradation models Biodegradation of organic carbon compounds is usually not constant (Janssen 1984). We tested three empirical models to predict biochar degradation, i.e. the first-order decay rate model, the double exponential model (Lehmann et al. 2009; Zimmerman et al. 2011), and a logarithmic model (Zimmerman 2010). All models facilitate curve simulation and half-life estimation, although with quite variable results as shown by the present study. Increasing model complexity often aims at interpreting results along the concept of multiple pools, e.g., total biochar degradation may involve one labile and one recalcitrant carbon pool. We applied RMSD as a first approximation for the goodness of fit of model data to measured data. As per its meaning, RMSD was able to compare the overall matching of different models by pointing to major mismatches between measured and model data. Over the period of 200 days, it appeared that the double exponential model followed by the first-order decay rate model were the best matches for the degradation curves. To assess the model performance, i.e., the goodness of fit weighed against the degree of parameterization (Burnham and Anderson 2004), we employed the corrected Akaike information criterion (AICC). The AICC showed overall preferences for the DEM over the FODM and for the FODM over the LOGM. However, the DEM was not always the best, and the difference in AICC between FODM and LOGM were sometimes small. Thus, the sum of results suggest that rather than calculating half-lives with only one model, an ensemble of models better captures the uncertainty inherent of empirical models. For a better model evaluation and improved process understanding, we need to investigate variously carbonized biochars and conduct more mechanistic studies on the early, midterm, and late degradation dynamics.
Acknowledgments Research contributing to this study was funded by the Deutsche Bundesstiftung Umwelt DBU. The authors are grateful to Rainer Georg Joergensen, University of Kassel, for his time invested to pre-review the manuscript. We thank Sonja Schimmelpfennig for sharing her analysis of biochar and feedstock pH and ash content, and Beate Lindenstruth and Ina Plesca for their technical support and discussion during the manuscript preparation.

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