Soil Biology & Biochemistry 97 (2016) 31e39

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Soil Biology & Biochemistry

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

Delayed and asymmetric responses of soil C pools and N uxes to

grassland/cropland conversions
E. Attard b, *, X. Le Roux a, X. Charrier c, O. Delfosse d, N. Guillaumaud a, G. Lemaire c,
S. Recous d, **
INRA, CNRS, Universit

e de Lyon, Laboratoire d'Ecologie Microbienne, UMR1418, UMR5557, 43 boulevard du 11 novembre 1918, F-69100 Villeurbanne,
France
b
Universit

e de Pau et des Pays de l'Adour, UMR5254 CNRS IPREM, F-64013 Pau, France
c
INRA, UE FERLUS, Les Verrines, F-86600 Lusignan, France
d
INRA, UMR614 FARE, 2 esplanade Roland Garros, F-51000 Reims, France
a

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 20 September 2015
Received in revised form
24 February 2016
Accepted 27 February 2016
Available online xxx

Agricultural lands are subject to changes in use for a variety of environmental and economic reasons.
Among these changes, shifts between crops and grasslands are particularly common. Understanding the
effects of these changes on soil functioning is crucial to anticipating crop productivity and environmental
impacts and yet has rarely been studied. The purpose of this study was to investigate the in situ temporal
dynamics of soil N cycling and C pools after grassland/cropland conversions and risks of N losses from
such systems. The experiment was conducted in western France at a long-term experimental site with a
temperate oceanic climate. We followed soil N cycling and C pools for four experimental treatments over
36 months: (i) continuous grassland, (ii) continuous rotation of annual crops, (iii) conversion from
grassland to a rotation of annual crops, and (iv) conversion from a rotation of annual crops to grassland.
Gross N mineralization, potential N immobilization and potential nitrication were estimated using the
15
N dilution/enrichment technique and FLUAZ model, based on samples from the 0e10 cm soil layer (0,
0.75, 3, 12, 24 and 36 months after change in land use) and 10e30 cm soil layer (after 24 and 36 months)
in all 4 treatments. Nitrogen pools (NH4, NO3, and organic N), total carbon (C), dissolved organic C,
microbial biomass C, and C mineralization were also determined on all soil samples.
The continuous grassland soil had rates of N mineralization and N immobilization twice as high as a
rotation of annual crops, as well as a slightly lower rate of nitrication and a higher organic C content.
Ploughing the grassland led to a rapid shift of soil organic matter pools, N uxes and microbial activities
towards characteristics of the cropland. These effects were attributed to cultivation mixing the soil layers.
In contrast, the restoration of grassland on soil previously cropped with annual species did not signicantly change soil N or C characteristics within 24 months although changes began to appear after 36
months. This time lag corresponded to the time needed for the complete establishment of root systems
for the grasses. Therefore we found a strong asymmetry in the patterns of change in gross N uxes and C
pools between the two conversions. Overall, we found that responses in N uxes were strongly correlated to soil C pools, except for nitrication. The ratio of potential nitrication to gross ammonium
immobilization, which indicates the risk of N losses in agricultural systems, was greater in annual crop
rotation than grassland. This indicates that adding grasslands into a rotation of annual crops does not
necessarily reduce the risk of N loss within 2 years but does reduce the risk after 3 years.
2016 Elsevier Ltd. All rights reserved.

Keywords:
15
N tracer technique
Mineralization-immobilization turnover
(MIT)
Nitrication
Soil organic carbon
Tillage

1. Introduction

* Corresponding author.
** Corresponding author.
E-mail addresses: eleonore.attard@univ-pau.fr (E. Attard), sylvie.recous@reims.
inra.fr (S. Recous).
http://dx.doi.org/10.1016/j.soilbio.2016.02.016
0038-0717/ 2016 Elsevier Ltd. All rights reserved.

Concerns about the effects of human activities on soil degradation, water quality, and greenhouse gas emissions are encouraging changes in agriculture systems including a shift towards
more sustainable land management. Among emerging practices,

32

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

farmers are reconsidering an old method of adding grasslands

within arable crop rotations, known as sod-based rotations or ley
farming. This type of farming may be a way to reconcile agricultural
productivity with concerns about environmental quality
(Franzluebbers et al., 2014; Lemaire et al., 2014) by integrating
crop-livestock systems (Russelle et al., 2007). These land-use shifts
have associated changes in plant cover and carbon (C) pools
(Matson, 1997; Balesdent et al., 2000) that can affect soil functioning, especially nitrogen (N) cycling that is crucial for maintaining soil quality, plant productivity and water quality. Soilvegetation interactions in permanent grasslands strongly couple
CeN dynamics, limiting the accumulation of mineral N, thus
reducing the risk of N losses to the atmosphere and hydrosphere
(Soussana and Lemaire, 2014). In an arable cropping system, these
plantesoil interactions are frequently interrupted, decoupling CeN
dynamics and increasing environmental risks of N loss (Drinkwater
and Snapp, 2007). Including grassland sequences within rotations
of arable crops may reduce these risks (Lemaire et al., 2015).
The use of the 15N isotope pool dilution/enrichment technique
has been intensively developed over the past 30 years to assess
mineralization-immobilization turnover (or MIT), under various
conditions. Studies have reported that microbial biomass and soil C
and N concentrations were highly correlated to gross N mineralization and immobilization (Barrett and Burke, 2000; Cookson et al.,
2007; Luxhi et al., 2008), while nitrication was correlated to N
availability and soil moisture (Attard et al., 2010; Simonin et al.,
2015). From a meta-analysis of 100 studies of soils under
different land uses and plant covers, Booth et al. (2005) concluded
that gross uxes of N mineralization and immobilization were
higher in grassland than cropland soils. Previous studies (e.g.,
Davidson et al., 1992; Hart et al., 1994; Recous et al., 1999; Niboyet
et al., 2011; Simonin et al., 2015) have shown that both net and
gross N mineralization and nitrication uxes are not necessarily
correlated, because different drivers affect gross mineralization,
gross immobilization and nitrication (Murphy et al., 2003, 2007).
For these reasons, measuring the underlying N cycling processes
and not just the resulting net N uxes may be crucial to understanding the effects of environmental disturbances on soil functioning (Schimel and Benett, 2004), particularly when these
disturbances affect the amount and nature of C inputs. Land uses
can have large impacts on C inputs, but few studies have reported
how they affect N uxes. Grassland establishment (Accoe et al.,
2004; Hamer et al., 2008), the conversion of fallow lands to croplands (Potthast et al., 2012), and changes in N fertilization (Hatch
et al., 2000) all affect gross N uxes in soils. However, these
studies compared agricultural systems that shifted agricultural
practices or land uses, several months or years after these changes
occurred. Such experiments do not reveal how fast changes
occurred and therefore limit our ability to anticipate how land use
conversions may affect the functioning of soils on time scales (crop
cycle to crop rotation) relevant to agricultural management. An
experimental design that includes ner-scale temporal monitoring
of soil characteristics can help address this issue. If the processes
underlying N dynamics respond differently to changes in land use, a
temporal imbalance in the N cycle could produce a period of
dysfunction in N cycling, such as a high nitrate accumulation
increasing the risk of N loss, or a loss of synchronization between
soil mineral N supply and plant demand.
Our aims were (i) to analyse the short- and medium-term effects
of converting grassland to annual crop rotations, and vice versa, on
the dynamics of N cycling processes and soil C pools. The results
were compared to control plots for which land use was unchanged,
i.e. continuous grassland or continuous rotation of annual crops;
and (ii) to analyse the kinetics of observed changes in different N
uxes for risks of N losses following land-use change. To do so, N

uxes and C and N pools were measured in parallel on soils

sampled 6 times over 3 years on plots under continuous grassland,
plots with continuous 3-year rotation of annual crops, plots converted from a 5-year old grassland to a rotation of annual crops, and
plots converted from a rotation of annual crops to a grassland.
2. Materials and methods
2.1. Experimental site, treatments and soil sampling
The experimental site used in this study has been described in
detail by Attard et al. (2011). The site is located in Lusignan, France
(46 240 51N, 0 070 14E) at the long-term observatory for environmental research (SOERE-ACBB, http://www.soere-acbb.com),
where the introduction of temporary grasslands into rotation is
being studied. The soil is a loamy textured Cambisol (FAO World
Reference Base for Soil Resources), that does not differ signicantly
between grassland and cropland plots (130 g clay kg1 soil,
690 g kg1 silt and 180 g kg1 sand). The climate is oceanic with
marked summer drought. The average monthly temperatures and
rainfall during the study period are presented as supplementary
material Fig. S1. We studied eight plots that had been continuously cropped for 5 years under a maize (Zea mays) e wheat (Triticum aestivum) e barley (Hordeum vulgare) rotation and eight plots
that had been grasslands for 5 years with Festuca arundinacea,
Dactylis glomerata and Lolium multiorum as the major plant species. Plots were 70 m  60 m for the rotation of annual crops and
20 m  12 m for the grassland. In March 2005, half of the plots (i.e.,
4 plots under annual crop rotation and 4 grassland plots) were
converted in land use. Half of the previous grassland plots were
ploughed to sow a maize crop, and half of the plots previously
cropped with barley were sown with a mixture of F. arundinacea,
D. glomerata and L. multiorum. This resulted in 4 treatments:
continuous rotation of annual crops (CC); continuous grassland
(GG); conversion from a 3-year rotation of annual crops to grassland (CG); and conversion from grassland to a 3-year rotation of
annual crops (GC). During the experiment, plots under the rotation
of annual crops (CC and GC) continued their maizeewheatebarley
rotation. The tillage that converted the grassland plots (GC treatment) incorporated 935 kg C ha1 and 50 kg N ha1 from the aerial
part of the grass plants and 1745 kg C ha1 and 55 kg N ha1 from
their roots into the top 25 cm of the soil. Tillage (25 cm depth) of
the CC and GC treatments occurred in October 2005, October 2006,
and May 2008. All major agricultural practices during the 36
months of the eld experiment and the initial C and N contents in
the 0e10 and 10e30 cm layers of grassland and cropland plots are
summarized in Table S1 (supplementary material).
The soil was sampled 6 times during the 36 months of the study,
i.e., just before the grassland/cropland conversion and 0.75, 3, 12,
24 and 36 months following conversion, except for the CC and GG
treatments, which were not sampled at 0.75 months. For each date,
ten soil cores were randomly sampled in the 0e10 cm layer of each
plot. Samples were taken from this layer because it is most likely to
gel-Knaber, 2005).
show immediate changes in SOM (Leifeld and Ko
Ten soil cores were also sampled in the 10e30 cm layer of each plot
at 24 and 36 months. For each plot and each soil layer, the ten cores
were pooled, and then passed through a 2 mm sieve before analysis.
The biomass of roots was sampled in the 0e30 cm layer once a
year for CC and CG soils and at T0 for GG soils. Soils were sampled in
triplicate in each plot. Roots were removed from soil with a 2 mm
sieve, washed with water to eliminate soil particles, and then dried
and weighed.
For each soil sample and each date, soil moisture (w/w) was
measured using a subsample (approximately 7 g) of each fresh soil
sample. The bulk density and total porosity of the upper soil layer

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

were measured in each plot on undisturbed soil using 215 cm3 steel
cylinders (one cylinder per plot) at the last sampling date. The
Water Filled Pore Space (WFPS), i.e. the fraction of total pore space
lled with water, was calculated according to Attard et al. (2011).
2.2. Measurements of gross N uxes
Measurements of N ux were quantied by the 15N pool dilution
technique (Murphy et al., 2003) using a soil amendment of
15
NeNH4. This approach allows simultaneous assessment of N
mineralization (also called ammonication), N immobilization and
nitrication. The measured ux of N mineralization is not modied
by 15NH4 supply (see, for instance, Luxhi et al., 2005); therefore, it
can be considered a gross activity, i.e., similar to actual gross N
mineralization without N addition. N immobilization and nitrication uxes were obtained during the same incubations; because
these uxes can be enhanced by the supply of 15NH4, they are
termed potential uxes hereafter.
For each sampling date and each treatment, a sub-sample of
fresh soil (60 g equivalent dry mass) was adjusted to 19% humidity
by spreading it thinly and spraying it with a 15(NH4)2SO4 solution
(15 mg NH4-N kg1 soil enriched at 5 atom% 15N) using an
atomizer. The soil was then mixed, separated into two sub-samples,
placed in glass vessels and incubated at 15 C. After 2 h, one vessel
was removed from the incubator and shaken (30 rotations min1)
with 100 mL of 1 M KCl for 30 min. After centrifugation (15 min;
5800g), the supernatants were ltered. The remaining pellet was
washed 3 times with 50 mL of 1 M KCl (shaking at 30 rotations
min1 for 15 min, centrifugation 5800g for 15 min) to dilute and
eliminate the residual labelled inorganic N (Recous et al., 1999). The
pellet was then dried for 24 h at 105 C and stored at ambient
temperature. The second sub-sample was removed from the incubator after 26 h and extracted in the same manner.
To measure the concentrations of N and abundance of 15N in the
KCl extracts using mass spectrometry, a microdiffusion technique
was used that sequentially transferred NH4 and NO3 to a lter
(Fillery and Recous, 2001, modied from Brooks et al., 1989). Briey,
MgO was added to the ask to transform NH4 into NH3 and trap it
in the form of (NH4)2SO4 on a lter acidied with H2SO4. After 7
days, the lter was removed, and a new one was added. Then,
Devarda's alloy was added to transform the NO3 into NH4. The
same procedure was used to trap the remaining mineral N on the
second lter. The NH4 and NO3 contents of the supernatants and
the organic N concentrations of the pellets, along with their 15N
abundances, were measured by mass spectrometry (N analyser
Euro EA, Eurovector, Milano, Italia and Delta Avantage, ThermoElectron, Bremen, Deutschland).
Gross N mineralization (m), potential N immobilization (i) (i.e.,
of NH4 (ia) plus NO3 (in)) and potential nitrication (n) were
calculated using the numerical model FLUAZ (Mary et al., 1998). The
model combines a numerical integration of differential equations
describing changes in N and 15N between four pools (NH4, NO3,
organic N and biomass N) after 2 and 26 h of incubation using a
nonlinear tting program. The model yields condence intervals of
estimates, correlation coefcients between tted parameters and a
criterion of the quality of t of predicted values with measured
values (MWE, mean weighted error). N mineralization and nitrication uxes were assumed to follow zero-order kinetics during
incubation. The parameter b, which is included in a Langmuir-type
equation and can vary from 0 (exclusive NH4 immobilization) to 1
(absence of preference of NH4 or NO3 in the N immobilization
process), was set to 0.05 according to Recous et al. (1999). For
clarity, abbreviations used in this study to identify gross N mineralization (m), total immobilization (i), immobilization from
ammonium (ia) and nitrication (n) calculated after addition of 15N

33

are those used in the FLUAZ (Mary et al., 1998).

2.3. Soil organic C, microbial biomass C and CeCO2 measurements
Total organic carbon concentration (TOC) was measured for
each sieved soil sample using an elemental analyser (NA 1500,
Carlo Erba, Milan, Italy). Microbial biomass carbon (MBC) was
determined using the fumigation-extraction method described by
Vance et al. (1987). The extraction of dissolved organic C (DOC)
from soil samples either fumigated with chloroform or not, was
performed with 40 mL of K2SO4 0.03 M (soil-solution 1:4, agitation
30 min, 20 C) and the concentration of DOC in extracts was analysed with a C-analyser (1010, O.I. analytical, Globalspec, NY, USA).
The amount of C in microbial biomass (MBC) was calculated as the
difference in DOC contents between fumigated and unfumigated
samples, with a correction factor KEC 0.38 (Vance et al., 1987).
C mineralization was estimated using CeCO2 emitted from soil
during the 24 h-incubations used to determine gross N uxes, and
was used as a proxy of soil heterotrophic microbial activity. Soil
samples were placed in jars containing a beaker with water and a
vial with 0.25 M of a NaOH solution to absorb CO2 produced. Sodium hydroxide contained in the vials was back titrated with
0.025 M HCl.
2.4. Statistical tests
For each treatment and each soil layer at a given date, each N
ux value was computed using the FLUAZ model (Mary et al., 1998),
as the average value for the 4 plots. The condence interval associated with each value was also computed by FLUAZ. Nonoverlapping condence intervals were considered to indicate signicant differences between treatments at each sampling date or
between sampling dates for each treatment.
Because we repeatedly sampled the same plots over 36 months,
we applied a two-way repeated measures ANOVA to test for effects
of treatment, sampling time and their interaction on TOC, DOC,
MBC, C mineralization and C:N. When signicant interactive effects
were observed, we performed a one-way ANOVA for each date to
determine treatment effects by date.
Pearson's test was used to evaluate correlations between N
uxes and soil environmental variables (WFPS, available NH4, TOC,
DOC, MBC, and emitted C mineralization). All statistical calculations
were performed with PAST software (Hammer et al., 2001).
3. Results
3.1. Dynamics of N uxes in the 0e10 cm soil layer
During the three-year experiment, gross N mineralization (m) in
the 0e10 cm layer was always signicantly higher in grasslands
(GG) than annual crops (CC); on average, it was two-fold higher in
GG (3.1 0.7 mg N kg1 soil day1) than CC soils (1.5 0.5 mg N kg1
soil day1) (Fig. 1a, Supplementary material Fig. S2). Potential N
immobilization (i) in the 0e10 cm layer followed the same trend and
was on average 2.9 0.9 mg N kg1 soil day1 for GG and
1.4 0.2 mg N kg1 soil day1 for CC (Fig. 1b, Supplemental material
Fig. S2). Both m and i in GG soils reached maximum values after 3
months (Fig. 1a, 1b), which corresponded to a severe drought period
(Supplementary material Fig. S3).
After conversion from grassland to cropland (i.e., 0.75 and 3
months after beginning the experiment), m and i in GC soils quickly
decreased and reached values similar to those observed in CC soils
(Fig. 1a,b); m and i remained similar between GC and CC treatments
at all other sampling dates. Conversely, conversion from annual
crops to grassland (CG) did not signicantly affect m and i during

34

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

at 36 months.
The ratio of potential nitrication to potential NH4 immobilization (n/ia) varied over time but was always higher in CC than GG
soils (average of 1.36 vs. 0.53, respectively) (Fig. 2), due mainly to
lower i uxes in the CC treatment (Fig. 1b). In the CG treatment, n/ia
was similar to the CC treatment during the rst 24 months, and
then decreased to values similar to the GG treatment at 36 months.
In the GC treatment, this ratio showed large uctuations over time
(due to concomitant changes in i and n) and reached a value close to
CC soils after 36 months (Fig. 2).
3.2. Vertical gradient of N uxes
Unlike the 0e10 cm layer, m and i in the 10e30 cm layer differed
markedly less between the four treatments at 24 and 36 months
(Fig. 3). As a consequence, vertical gradients of m and i uxes were
more pronounced for grassland treatments, i.e., GG and CG soils,
compared to CC and GC treatments (Fig. 3). n was also lower in the
10e30 cm than 0e10 cm soil layers for the GG, GC, CC soils, while it
was similar for the 2 layers in CG soils (data not shown).
3.3. Dynamics of C in the 0e10 cm soil layer
Total organic carbon (TOC), microbial biomass carbon (MBC) and
CeCO2 emitted (C mineralization) from the 0e10 cm soil layer, were
signicantly higher in GG than CC soils over the entire period
(Table 1). On average, these values were approximately twice as
high in GG than CC soils, with TOC concentrations of 17 and
10 g C kg1 soil, MBC concentrations of 561 and 287 mg C kg1 soil,
and C mineralization rates of 27.3 and 14.5 mg C kg1 soil day1,
respectively. Dissolved Organic Carbon (DOC) concentrations were
also higher in GG than CC soils throughout the experiment, but
these differences were not signicant (Table 1).
Conversion from grassland to cropland (GC treatment) signicantly decreased TOC, MBC and C mineralization, 0.75 months after
conversion (Table 1). TOC in GC soils remained at intermediate
values between those observed for control treatments, GG and CC
soils, during the entire experiment. The conversion from annual
crops to grassland (CG treatment) did not signicantly affect TOC,
MBC and C mineralization, although there was a tendency for C
pools to increase by 36 months (Table 1).
Considering all treatments and soil layers, TOC, MBC, DOC and C
mineralization were positively and signicantly correlated (TOC vs.
DOC R2 0.70 p < 0.01; TOC vs. MBC R2 0.70 p < 0.01; TOC vs. C
3.0

the rst 24 months (Fig. 1a,b). However, after 36 months, m and i in

CG soils increased to values similar to GG soils (Fig. 1a,b).
Potential nitrication (n) in the 0e10 cm layer did not differ
initially between GG and CC treatments, but after 12 months, n was
higher in CC than GG soils (Fig. 1c). n was similar in both CG and GG
soils throughout the experiment at approximately 1.5 mg N kg1
soil day1 (Fig. 1c, Supplementary material Fig. S2). In GC soils, n
remained low compared to values in the other treatments during
the rst 24 months but then increased to values similar to CC soils

2.5
2.0

Ratio n / ia

Fig. 1. Temporal dynamics of gross N uxes in the 0e10 cm soil layer under different
types of land use. (A) gross N mineralization; (B) potential N immobilization; (C) potential nitrication. C: continuous grasslands (GG); B: conversion from grassland to
cropland (GC); :: continuous rotation of annual crops (CC); : conversion from
cropland to grassland (CG). Mean values of uxes (n 4) calculated by the FLUAZ
model. Bars are condence intervals of the means. The condence intervals that are
not visible have a smaller size than that of the symbols.

1.5
1.0
0.5

0.0
0

12

18

24

30

36

Time (months)

Fig. 2. Ratio of potential nitrication to potential NH4 immobilization (n/ia). C:

continuous grasslands (GG); B: conversion from grassland to cropland (GC); ::
continuous rotation of annual crops (CC); : conversion from annual crops to grassland (CG). Mean values of uxes (n 4 replicates) calculated by the FLUAZ model.

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

N mineralization

35

mg N kg -1 soil day-1

N immobilization

010cm
1030cm

24 36
GG

24 36
GC

24 36
CG

24 36
CC

24 36
GG

24 36
GC

24 36
CC

24 36
CG

Fig. 3. Gross N mineralization and potential N immobilization observed 24 and 36 months after changes in land use, in the 0e10 cm and 10e30 cm soil layers. Mean values (n 4
replicates) and associated condence intervals are calculated by the FLUAZ model.

Table 1
Total organic carbon concentration (TOC), microbial biomass content (MBC), C mineralization, dissolved organic carbon concentration (DOC) and C:N ratio of the bulk soil in the
0e10 cm soil layer for the 4 treatments (continuous grasslands GG; continuous cropland (rotation of annual crops) CC; conversion from grassland to cropland GC; and
conversion from cropland to grassland CG). Mean values (n 4 replicates). For each sampling date, different letters indicate signicant difference between treatments at
p < 0.05. nd not determined.
Treatment

Time after change in land use (months)

0.75

12

24

TOC (g kg1)

GG
CC
GC
CG

0
17.7
11.4
18.1
10.7

a
b
a
b

16.2 a
nd
14.6 b
nd

15.2 a
10.0 c
12.8 b
9.6 c

16.4 a
9.9 c
14.3 b
10.1 c

18.6
10.1
13.4
10.1

a
c
b
c

18.2 a
9.9 c
13.3 b
11.5 c

MBC (mg kg1)

GG
CC
GC
CG

525
301
562
297

a
b
a
b

469 a
nd
362 b
nd

411
205
264
199

a
bc
b
c

546
284
391
315

a
b
b
b

564
349
278
310

a
b
b
b

853
296
361
408

a
b
b
b

C mineralization (mg kg1 d1)

GG
CC
GC
CG

24.0
11.3
27.4
11.5

a
b
a
b

22.5 a
nd
12.4 b
nd

37.8
12.7
12.0
12.9

a
b
b
b

21.4
12.4
15.8
15.5

ns
ns
ns
ns

21.6
13.0
16.7
17.8

a
b
a
a

36.7
19.4
18.8
25.2

a
b
b
b

DOC (mg kg1)

GG
CC
GC
CG

37.0
15.2
35.7
14.8

ns
ns
ns
ns

24.7 ns
nd
23.7 ns
nd

25.3
10.7
21.3
10.7

ns
ns
ns
ns

39.3
18.0
24.1
19.6

ns
ns
ns
ns

23.4
11.7
19.2
10.5

ns
ns
ns
ns

26.2 ns
7.4 ns
15.3 ns
9.1 ns

Soil C:N

GG
CC
GC
CG

9.7
8.8
9.9
8.9

9.8 ns
nd
9.6 ns
nd

10.4 ns
9.6 ns
10.0 ns
9.7 ns

10.0 ns
9.1 ns
9.6 ns
9.3 ns

11.3
10.2
10.6
10.3

ns
ns
ns
ns

11.7
10.2
10.6
10.6

ns
ns
ns
ns

mineralization R2 0.39 p < 0.01; DOC vs. MBC R2 0.43 p < 0.01;
DOC vs. C mineralization R2 0.18 p < 0.05; MBC vs. C mineralization R2 0.59 p < 0.01).
After conversion from annual crops to grassland, the C content
of roots in CG soils increased and rapidly exceeded values observed
in CC soils (Fig. 4). At the end of the experiment, root C in CG soils
was 15-fold higher than CC soils. The accumulation of root biomass
(and C) in CG soils was progressive, representing 30% and 80% of the
amounts in GG soils after 12 and 24 months, respectively, and
reaching similar to or even higher values than GG soils by 36
months (Fig. 4).

36

ns
ns
ns
ns

at 24 and 36 months (Fig. 5aec). By contrast, in CC soils that were

tilled each year, TOC and DOC did not differ between soil layers,
although C mineralization was signicantly different between
layers at both dates. For the GC treatment, differences observed in
TOC, DOC and C mineralization between the 0e10 and 10e30 cm
soil layers were signicant or marginally signicant, but the
amplitude of the vertical gradients was weak (Fig. 5aec). The CG
treatment followed a pattern similar to the CC treatment, except
that a slight but signicant gradient appeared at 36 months for both
TOC and DOC (Fig. 5aec).
3.5. Relationships between N uxes and C in soils

3.4. Vertical gradient of C in the soil

TOC and DOC concentrations and C mineralization in GG soils
were signicantly lower in the 10e30 cm than 0e10 cm soil layers

Signicant, positive correlations were observed between

changes in m and organic C pools. TOC was the best predictor of m
in the 0e10 cm soil layer for all treatments at all dates (R2 0.63,

36

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

Fig. 4. Dynamics of root biomass carbon in the 0e30 cm soil layer after grassland
establishment on plots previously grown with annual crops (CG treatment) and
comparison to root biomass C in continuous grassland (GG) and annual crops (CC) at
the beginning of the experiment (dashed lines).

p < 0.01; Table 2 and Fig. 6), but changes in m were also correlated
with changes in MBC (R2 0.41 p < 0.01) and C mineralization
(R2 0.56 p < 0.01). i was strongly and positively correlated to m
(R2 0.84 p < 0.01) (Table 2). Other CeN relationships were
examined among treatments, such as the C:N ratio of bulk soil, and
the ratios of gross mineralization:TOC and gross N mineralization:C
mineralization, but no signicant differences between treatments
were found. There were also no signicant differences in the C:N
ratio of the 10e30 cm soil layer after 24 and 36 months.
No single relationship was found between n and either soil NH4
concentration or WFPS (data not shown). Changes in n in CC and CG
treatments on one hand and GG treatment on the other were
correlated to changes in WFPS (Fig. 7; for CC and CG R2 0.48
p < 0.05, for GG R2 0.91 p < 0.05). These relationships for the GC
treatment were not signicant.

4. Discussion
4.1. N uxes and C pools in soils under rotation of annual crops vs.
continuous grassland
Agricultural management signicantly affected N uxes in the
upper soil layer, with both gross N mineralization and potential
immobilization being higher in continuous grassland than in
annual crop soils throughout the 3 years of this experiment. As
reported previously by Silva et al. (2005) and Muruganandam et al.
(2010), these differences in rates of gross N uxes are caused by
faster microbial turnover driven by greater root C supply and
organic matter accumulation in the top layer of soil in grasslands
than croplands. Consequently, in the lower soil layers where root
growth and C deposition are lower, differences in N mineralization
and immobilization uxes were much lower between grasslands
and croplands. This explanation is consistent with higher concentrations of total C, dissolved organic C, microbial biomass C and
higher rate of C mineralization in the 0e10 cm layer of grassland
soils compared to annual crop soils, and with sharper vertical
gradients observed for these pools in grassland soils. This pattern is
generally reported for grasslands (Schimel et al., 1989).
Three months after beginning the experiment, we observed a
large increase in gross N mineralization and immobilization uxes
in grassland soils. We assume that this one-time increase could
have been a synergistic effect of microbial mortality during the
pronounced summer drought followed by rewetting the soil in the
laboratory prior to 15N dilution, resulting in high N uxes. This

Fig. 5. Total organic carbon concentration (TOC), dissolved organic carbon concentration (DOC) and carbon mineralization at 24 and 36 months after changes in land use
for the 0e10 cm and 10e30 cm soil layers (n 4 replicates, bars are standard errors). *
indicates signicant differences between soil layers for a given treatment and a given
date at p < 0.05; # indicates signicant differences at p < 0.10; ns means not signicantly different.

effect has been reported by Dijkstra et al. (2012) for N uxes within
a few days after the addition of water. Fierer and Schimel (2003)
proposed that rewetting a dry soil led to cell lysis and aggregate
disruption and a release of labile organic matter available to microorganisms for C mineralization.
Cropland soils had higher rates of potential nitrication than
grassland soils. This difference was previously reported by Li and
Lang (2014) for gross nitrication rate in cultivated vs. uncultivated soils. Potential nitrication ux was not correlated with soil
C, and so there was no relationship between nitrication and gross
mineralization or immobilization. Our results showed that

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

37

Table 2
Single factor linear regression analysis between N uxes, C mineralization rates and selected explanatory variables for both soil layers for the 4 treatments and all sampling
dates. For the N uxes, the regression was calculated without and with (value in brackets) the GG treatment at 3 months after the start of the experiment (corresponding to the
drought event). ** indicates signicant correlation at p < 0.01. ns: correlation not signicant (p > 0.05). Units of each measurement are the same as in Fig. 1 and Table 1.
Gross N Mineralization

Explanatory variable
TOC
MBC
DOC
NO3 Concentration
NH4 Concentration
Soil C/N
Gross N Mineralization
Potential N Immobilization
Potential Nitrication

Potential
Nitrication

C Mineralization

R2

R2

R2

R2

0.63 (0.57)
0.51 (0.41)

**
**
ns
ns
ns
ns

0.59 (0.50)
0.47 (0.33)

**
**
ns
ns
ns
ns
**

e
e
e
e
e
e
e
e

ns
ns
ns
ns
ns
ns
ns
ns

0.40
0.57

**
**
ns
ns
**
ns
**
**
ns

e
e
e

e
e
e
0.84

e
0.32
0.56
0.65
e

Gross mineralization

5.0

mg N kg-1 soil day-1

Potential N
Immobilization

y = 0.20x - 0.69
R2 = 0.63
p <0.01

4.0
3.0
2.0
1.0
0.0

10

12

14

16

18

20

TOC (g C kg-1 soil)

Fig. 6. Relationship between gross N mineralization and total soil carbon (TOC) in soil.
Each point corresponds to a mean value of a treatment at a given date (n 4 replicates) for the 0e10 cm soil layer. C: continuous grasslands (GG); B: conversion from
grassland to rotation of annual crops (GC); :: continuous rotation of annual crops
(CC); : conversion from annual crops to grassland (CG). The symbols in gray
correspond to the 10e30 cm layer at 24 and 36 months.

nitrication was correlated with WFPS, the relative role of this

factor differing between GG and CC soils. GG soils were indeed
characterized by a relatively high availability of ammonium (due to
high m ux) and low WFPS (restricting the n ux), likely due to
higher transpiration by vegetation. In contrast, CC soils tended to
have higher WFPS. Le Roux et al. (2008) and Gleeson et al. (2010)
showed that environmental drivers, like soil moisture and N
availability, effected temporal changes in nitrication directly or
indirectly via changes in community structure and abundance of
ammonia oxidizers over periods of a few weeks or months.
4.2. Responses of C and N pools and uxes to grassland-cropland
conversions
The conversion from grassland to cropland (GC) led to a significant decrease in all C pools; by 0.75 months for TOC and MBC, and
12 months for DOC. We explain these rapid changes as a result of
soil tillage enhancing organic matter mineralization and diluting
the upper layer by mixing it with deeper soil. Along with many
other authors (Balesdent et al., 1998; Post and Kwon, 2000;
Butterbach-Bahl and Gundersen, 2011), we attribute this fast dynamic to the initial mechanical disruption of the grassland soil

Fig. 7. Relationships between potential nitrication and Water Filled Pore Space.
Symbols are as in Fig. 5. Each point corresponds to a mean value for one treatment at
one sampling date (n 4 replicates). Data for the sampling at 3 months were excluded
due to a severe drought.

prole that increased the vulnerability of SOM under cultivation.

These initial changes were maintained by annual soil tillage and
reduced C inputs by annual crops compared to perennial grasses.
However, both TOC and DOC in GC plots stayed at intermediate
levels in the 0e10 cm layer between those observed in the
continuous annual crops (CC) and continuous grassland soils (GG).
This suggests that C inherited from the previous grassland was
relatively stable over 3 years. This observation is consistent with a
previous study on the same site (Rumpel and Chabbi, 2010), which
reported that soil organic matter in GC plots retained some characteristics typical of grassland management, e.g., non-cellulosic
carbohydrates were preserved for at least 2 years. However, both
microbial biomass C and respiration responded more quickly to
changed agricultural practices than TOC or DOC, probably due to
more rapid turnover and because microbial biomass responds to C
inputs from litter and root exudates. It is notable that tilling the
grassland incorporated approximately 7 Mg ha1 of plant biomass
in soils but did not increase gross N mineralization. A fast degradation of plant residues with high C quality and N concentrations
(e.g., Sanaullah et al., 2010) is the likely fate for grassland biomass,

38

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

and we believe that most mineralization occurred before the rst

sampling at 0.75 months following grassland tillage. This hypothesis is supported by Recous et al. (1999) and Shindo and Nishio
(2005) who showed an increase in gross N mineralization and
immobilization 7 days after the incorporation of wheat straw, followed by a large decrease in these uxes after only one month.
Conversely, the conversion from rotation of annual crops to
grassland (CG) did not signicantly affect any C pools or uxes
before 3 years. These variables were strikingly stable over the rst
24 months, and they started to increase only at the last sampling
date. This nding agrees with other studies showing that several
years are needed for a signicant increase in soil C storage after
sowing grasslands. For example, C pools were much lower in 6 and
14-year old grasslands than in a 50-year old grassland (Accoe et al.,
2004). This observation also matches the progressive increase in
root biomass that we observed in the newly established grassland,
suggesting that cumulative C inputs from grassland vegetation to
soils are not linearly related to time. Actually, the root system develops rst, which increasingly contributes to C accumulation in
soil via rhizodeposition and root senescence. This gradual establishment of root systems could have increased gross N uxes
observed after 3 years, via a rhizosphere priming effect, even
though no signicant increase in total C was observed. Zhu et al.
(2014) observed a concomitant increase in C mineralization and
gross N mineralization in soils with more living roots. Similarly,
Bengtson et al. (2012) observed a strong correlation between gross
N mineralization and root exudates.
Consequently, we observed a strong asymmetry in effects of
land-use changes between grassland and cropland. Our results
clearly showed a time lag of at least 3 years before signicant effects
of the conversion from annual crops to grassland altered soil C and
MIT. Conversely, the conversion of grassland to annual crops showed
fast changes to soil C dynamics (Post and Kwon, 2000; McLauchlan,
2006). This asymmetry in the evolution of soil characteristics between the two systems shows the importance of studying long-term
effects of incorporating temporary grasslands in crop rotation on
both soil fertility and environmental impacts. While the whole
system approach of the SOERE ACBB eld experiment does not
permit disentangling the effects of individual controlling factors,
high correlations between gross N mineralization and immobilization, and C dynamics are consistent with previous works (Recous
et al., 1999; Cookson et al., 2007; Luxhi et al., 2008). In this study,
grassland/cropland conversions had not actually altered relationships between C and N dynamics; instead, we conrmed strong
connections between changes in N uxes and soil C.
4.3. Potential implications for soil N availability and N losses
One of the most important concerns for sustainable management of agricultural systems is the loss of N, primarily through NO3
leaching and denitrication. The ratio of Nitrication/Ammonium
Immobilization (n/ia) is frequently used to assess the risk of N
leaching (Stockdale et al., 2002). Its value indicates the level of
competition for ammonium between all microbiota vs. nitriers,
i.e., the balance between ammonium storage in microbial biomass
and oxidation of ammonium to nitrate by nitriers. Several authors
have shown a higher n/ia ratio for cropped soils than grasslands
(e.g., 1.86 vs. 0.61, respectively, Booth et al., 2005). Our results are
consistent with this report, with n/ia ratios of 1.36 and 0.53 for
crops and grasslands, respectively. In our grasslands, NH4 was
preferentially consumed and stored in microbial biomass probably
due to high C availability (low n/ia ratio), diminishing the risk of N
leaching and/or denitrication. In contrast, nitriers preferentially
used NH4 in soils under rotation of annual crops (high n/ia ratio),
likely increasing the probability of N leaching and/or

denitrication. This notion is supported by previous measurements

of NO
3 leaching under continuous grassland and continuous
cropland for 3 years on our experimental site (Kunrath et al., 2015).
These authors showed that nitrate concentrations in drainage water averages 57 mg L1 from cropland and only 10 mg L1 from
grassland despite high N fertilization of both. In conversion from
grassland to cropland, the n/ia ratio remained low for most of the
rst 24 months. Similarly, the denitrication potential of the same
soils also stayed low (Attard et al., 2011). The benet of grasslands
in terms of a more conservative N cycle in the soil could last a few
years after tilling the grassland. Again, this is supported by measurements of NO
3 leaching after the conversion of a 6-year grassland to rotation of annual crops on our experimental site (Kunrath
et al., 2015), which showed no increased loss of N, compared to
intact grassland. During the rst 24 months following the conversion of cropland to grassland, the n/ia ratio remained high and
similar to the n/ia for continuous rotation of annual crops (mainly
due to low immobilization rates), decreasing only after 36 months.
In terms of agricultural management, establishing grassland on a
previously cropped soil does not necessarily reduce the risk of N
loss over the medium term (here 2 years) and so maintaining
grasslands over sufciently long periods is necessary to conserve
soil N cycling.
Acknowledgements
The project was funded by the GESSOL2 programme A02544 of
the French Ministry of Environment and Sustainable Development
(MEDD), and by the ECCO-PNBC programme (COSMOS-Flux proj
gion
ect). The PhD of E. Attard was funded by INRA and the Re
Poitou-Charentes. The authors thank Eric Grehan and Gonzague
Alavoine (INRA) for help during eld sampling and microdiffusion,
and the staff of the Observatory for Environmental Research Agroecosystem, biogeochemical cycles and biodiversity (SOERE-ACBB)
in Lusignan. We thank two anonymous reviewers for their
insightful comments and suggestions, and Professor Daryl Moorhead for his careful reading of the nal version of this manuscript.
Appendix A. Supplementary data
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.soilbio.2016.02.016.
References
Accoe, F., Boeckx, P., Busschaert, J., Hofman, G., Van Cleemput, O., 2004. Gross N
transformation rates and net N mineralisation rates related to the C and N
contents of soil organic matter fractions in grassland soils of different age. Soil
Biol. Biochem. 36, 2075e2087.
Attard, E., Poly, F., Laurent, F., Commeaux, C., Terada, A., Smets, B., Recous, S., Le
Roux, X., 2010. Shifts between Nitrospira- and Nitrobacter-like nitrite oxidizers
underly the response of soil nitrite oxidizing enzyme activity to changes in
tillage practices. Environ. Microbiol. 12, 315e326.
Attard, E., Recous, S., Chabbi, A., De Berranger, C., Guillaumaud, N., Labreuche, J.,
Philippot, L., Schmid, B., Le Roux, X., 2011. Soil environmental conditions rather
than denitrier abundance and diversity drive potential denitrication after
changes in land uses. Global Change Biol. 17, 1975e1989.
Balesdent, J., Besnard, E., Arrouays, D., Chenu, C., 1998. The dynamics of carbon in
particle-size fractions of soil in a forest-cultivation sequence. Plant Soil 201,
49e57.
Balesdent, J., Chenu, C., Balabane, M., 2000. Relationship of soil organic matter
dynamics to physical protection and tillage. Soil Tillage Res. 53, 215e230.
Barrett, J.E., Burke, I.C., 2000. Potential nitrogen immobilization in grassland soils
across a soil organic matter gradient. Soil Biol. Biochem. 32, 1707e1716.
Bengtson, P., Barker, J., Grayston, S.J., 2012. Evidence of a strong coupling between
root exudation, C and N availability, and stimulated SOM decomposition caused
by rhizosphere priming effects. Ecol. Evol. 2, 1843e1852.
Booth, M.S., Stark, J.M., Rastetter, E., 2005. Controls on nitrogen cycling in terrestrial
ecosystems: a synthetic analysis of literature data. Ecol. Monogr. 75, 139e157.
Brooks, P.D., Stark, J.M., McInteer, B.B., Preston, T., 1989. Diffusion method to prepare
soil extracts for automated N-15 analysis. Soil Sci. Soc. Am. J. 53, 1707e1711.

E. Attard et al. / Soil Biology & Biochemistry 97 (2016) 31e39

Butterbach-Bahl, K., Gundersen, P., 2011. Nitrogen processes in terrestrial ecosystems. In: Sutton, M.A., Howard, C.M., Erisman, J.W., Billen, G., Bleeker, A.,
Grennfelt, P., van Grinsven, H., Grizzetti, B. (Eds.), The European Nitrogen
Assessment. Cambridge University Press, pp. 99e125 (Chapter 6).
Cookson, W.R., Osman, M., Marschner, P., Abaye, D.A., Clark, I., Murphy, D.V.,
Stockdale, E.A., Watson, C.A., 2007. Controls on soil nitrogen cycling and microbial community composition across land use and incubation temperature.
Soil Biol. Biochem. 39, 744e756.
Davidson, E.A., Hart, S.C., Firestone, M.K., 1992. Internal cycling of nitrate in soils of a
mature coniferous forest. Ecology 73, 1148e1156.
Dijkstra, F.A., Augustine, D.J., Brewer, P., von Fischer, J.C., 2012. Nitrogen cycling and
water pulses in semiarid grasslands: are microbial and plant processes
temporally asynchronous? Oecologia 170, 799e808.
Drinkwater, L.E., Snapp, S.S., 2007. Nutrients in agroecosystems: rethinking the
management paradigm. Adv. Agron. 92, 163e186.
Fierer, N., Schimel, J.P., 2003. A proposed mechanism for the pulse in carbon dioxide
production commonly observed following the rapid rewetting of a dry soil. Soil
Sci. Soc. Am. J. 67, 798e805.
Fillery, I.R.P., Recous, S., 2001. Use of enriched 15N sources to study soil N transformations. In: Unkovich, M., Pate, J.S., McNeill, A.M., Gibbs, D. (Eds.), Stable
Isotope Techniques in the Study of Biological Processes and Functioning of
Ecosystems. Kluwer Academic, Dordrecht, the Netherlands, pp. 167e194.
Franzluebbers, A.J., Sawchik, J., Taboada, M., 2014. Agronomic and environmental
impacts of pasture crop rotations in temperate North and South America. Agric.
Ecosyst. Environ. 190, 18e26.
Gleeson, D.B., Mller, C., Banerjee, S., Ma, W., Siciliano, S.D., Murphy, D.V., 2010.
Response of ammonia oxidizing archaea and bacteria to changing water lled
pore space. Soil Biol. Biochem. 42, 1888e1891.
Hamer, U., Makeschin, F., Stadler, J., Klotz, S., 2008. Soil organic matter and microbial
community structure in set-aside and intensively managed arable soils in NESaxony, Germany. Appl. Soil Ecol. 40, 465e475.
Hammer, ., Harper, D.A.T., Ryan, P.D., 2001. PAST: paleontological statistics software package for education and data analysis. Palaeontol. Electron. 4, 9.
Hart, S.C., Nason, G.E., Myrold, D.D., Perry, D.A., 1994. Dynamics of gross nitrogen
transformations in an old-growth forest e the carbon connection. Ecology 75,
880e891.
Hatch, D.J., Lovell, R.D., Antil, R.S., Jarvis, S.C., Owen, P.M., 2000. Nitrogen mineralization and microbial activity in permanent pastures amended with nitrogen
fertilizer or dung. Biol. Fertil. Soils 30, 288e293.

ranger, C., Charrier, X., Gastal, F., de Facio Carvalho, P.C.,
Kunrath, T.R., de Be
Lemaire, G., Emile, J.-C., Durand, J.-L., 2015. How much do sod-based rotations
reduce nitrate leaching in a cereal cropping system? Agric. Water Manag. 150,
46e56.
Le Roux, X., Poly, F., Currey, P., Commeaux, C., Hai, B., Nicol, G.W., Prosser, J.I.,
Schloter, M., Attard, E., Klumpp, K., 2008. Effects of aboveground grazing on
coupling among nitrier activity, abundance and community structure. ISME J.
2, 221e232.
gel-Knaber, I., 2005. Soil organic matter fractions as early indicators for
Leifeld, J., Ko
carbon stock changes under different land use? Geoderma 124, 143e155.
Lemaire, G., Franzluebbers, A.J., Carvalho, P.C., Dedieu, B., 2014. Integrated croplivestock systems: strategies to achieve synergy between agricultural production and environmental quality. Agric. Ecosyst. Environ. 190, 4e8.
Lemaire, G., Gastal, F., Franzluebbers, A.J., Chabbi, A., 2015. Grassland-cropping rotations: an avenue for agricultural diversication to reconcile high production
with environmental quality. Environ. Manag. 56, 1065e1077.
Li, P., Lang, M., 2014. Gross nitrogen transformations and related N2O emissions in
uncultivated and cultivated black soil. Biol. Fertil. Soils 50, 197e206.
Luxhi, J., Fillery, I.R.P., Murphy, D.V., Bruun, S., Jensen, L.S., Recous, S., 2008. Distribution and controls on gross N mineralization-immobilization-turnover in
soil subjected to zero tillage. Eur. J. Soil Sci. 59, 190e197.
Luxhi, J., Recous, S., Fillery, I.R.P., Murphy, D.V., Jensen, L.S., 2005. Comparison of
(NH4)-N-15 pool dilution techniques to measure gross N uxes in a coarse
textured soil. Soil Biol. Biochem. 37, 569e572.

39

Mary, B., Recous, S., Robin, D., 1998. A model for calculating nitrogen uxes in soil
using 15N tracing. Soil Biol. Biochem. 30, 1963e1979.
Matson, P.A., 1997. Agricultural intensication and ecosystem properties. Science
277, 504e509.
Mclauchlan, K.K., 2006. The nature and longevity of agricultural impacts on soil
carbon and nutrients: a review. Ecosystems 9, 1364e1382.
Murphy, D.V., Recous, S., Stockdale, E.A., Fillery, I.R.P., Jensen, L.S., Hatch, D.J.,
Goulding, K.W.T., 2003. Gross nitrogen uxes in soil: theory, measurement and
application of 15N pool dilution techniques. Adv. Agron. 79, 69e118.
Murphy, D.V., Stockdale, E.A., Poulton, P.R., Willison, T.W., Goulding, K.W.T., 2007.
Seasonal dynamics of carbon and nitrogen pools and uxes under continuous
arable and ley-arable rotations in a temperate environment. Eur. J. Soil Sci. 58,
1410e1424.
Muruganandam, S., Israel, D.W., Robarge, W.P., 2010. Nitrogen transformations and
microbial communities in soil aggregates from three tillage systems. Soil Sci.
Soc. Am. J. 74, 120e129.
Niboyet, A., Le Roux, X., Dijkstra, P., Hungate, B.A., Barthes, L., Blankinship, J.C.,
Brown, J.R., Field, C.B., Leadley, P.W., 2011. Testing interactive effects of global
environmental changes on soil nitrogen cycling. Ecosphere 2. http://dx.doi.org/
10.1890/ES10-00148.1. Article 56.
Post, W.M., Kwon, K.C., 2000. Soil carbon sequestration and land-use change:
processes and potential. Global Change Biol. 6, 317e328.
Potthast, K., Hamer, U., Makeschin, F., 2012. Land-use change in a tropical mountain
rainforest region of southern Ecuador affects soil microorganisms and nutrient
cycling. Biogeochemistry 111, 151e167.
Recous, S., Aita, C., Mary, B., 1999. In situ changes in gross N transformations in bare
soil after addition of straw. Soil Biol. Biochem. 31, 119e133.
Rumpel, C., Chabbi, A., 2010. Response of bulk chemical composition, lignin and
carbohydrate signature to grassland conversion in a ley-arable cropping system.
Nutr. Cycl. Agroecosyst. 88, 173e182.
Russelle, M.P., Entz, M.H., Franzluebbers, A.J., 2007. Reconsidering integrated croplivestock systems in North America. Agron. J. 99, 325e334.
Sanaullah, M., Chabbi, A., Lemaire, G., Charrier, X., Rumpel, C., 2010. How does plant
leaf senescence of grassland species inuence decomposition kinetics and litter
compounds dynamics? Nutr. Cycl. Agroecosyst. 88, 159e171.
Schimel, J.P., Benett, J., 2004. Nitrogen mineralization: challenges of a changing
paradigm. Ecology 85, 591e602.
Schimel, J.P., Jackson, L.E., Firestone, M.K., 1989. Spatial and temporal effects of plant
microbial competition for inorganic nitrogen in a California annual grassland.
Soil Biol. Biochem. 21, 1059e1066.
Shindo, H., Nishio, T., 2005. Immobilization and remineralization of N following
addition of wheat straw into soil: determination of gross N transformation rates
by N-15-ammonium isotope dilution technique. Soil Biol. Biochem. 37,
425e432.
Silva, R.G., Jorgensen, E.E., Holub, S.M., Gonsoulin, M.E., 2005. Relationships between culturable soil microbial populations and gross nitrogen transformation
processes in a clay loam soil across ecosystems. Nutr. Cycl. Agroecosyst. 71,
259e270.
Simonin, M., Le Roux, X., Poly, F., Lerondelle, C., Hungate, B.A., Nunan, N., Niboyet, A.,
2015. Coupling between and among ammonia-oxidizers and nitrite-oxidizers in
grassland mesocosms submitted to elevated CO2 and nitrogen supply. Microb.
Ecol. 70, 809e818.
Soussana, J.-F., Lemaire, G., 2014. Coupling carbon and nitrogen cycles for environmentally sustainable intensication of grasslands and crop-livestock systems. Agric. Ecosyst. Environ. 190, 9e17.
Stockdale, E.A., Hatch, D.J., Murphy, D.V., Ledgard, S.F., Watson, C.J., 2002. Verifying
the nitrication to immobilisation ratio (N/I) as a key determinant of potential
nitrate loss in grassland and arable soils. Agronomie 22, 831e838.
Vance, E.D., Brookes, P.C., Jenkinson, D.S., 1987. An extraction method for measuring
soil microbial biomass-C. Soil Biol. Biochem. 19, 703e707.
Zhu, B., Gutknecht, J.L.M., Herman, D.J., Keck, D.C., Firestone, M.K., Cheng, W., 2014.
Rhizosphere priming effects on soil carbon and nitrogen mineralization. Soil
Biol. Biochem. 76, 183e192.