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Applied Soil Ecology

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

Soil organic carbon, nutrients and relevant enzyme activities in particle-size

fractions under conservational versus traditional agricultural management
Shuping Qin a,b , Chunsheng Hu a,∗ , Xinhua He c , Wenxu Dong a , Junfang Cui a,b , Ying Wang a,b
a
Key Laboratory of Agricultural Water Resources, Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology,
The Chinese Academy of Sciences, 286 Huaizhong Road, Shijiazhuang, Hebei 050021, China
b
Graduate University of the Chinese Academy of Sciences, Beijing 100049, China
c
Northern Research Station, USDA Forest Service, Houghton, MI 49931, USA

a r t i c l e i n f o a b s t r a c t

Article history: Micro-scale investigation is helpful for better understanding of the relationships between organic mat-
Received 3 November 2009 ter, microorganisms and nutrients in soil, and for better interpretation of modifications induced by soil
Received in revised form 14 March 2010 management. The soil particle-size fractions (2000–200, 200–63, 63–2, and 2–0.1 ␮m), contents of soil
Accepted 16 March 2010
organic carbon (SOC), total N (STN), available P (SAP), dissolved organic C (DOC), light fraction organic C
(LOC), microbial biomass N (MBN), basal respiration (SBR), and relevant enzyme activities of C, N and P
Keywords:
transformations, such as ␤-glucosidase (␤-G), N-acetyl-␤-glucosaminidase (N-G), protease, urease and
Conservational agriculture
alkaline phosphomonoesterase (APH) were analyzed to study the effects of 8-year-period conservational
Particle-size fractions
Soil enzyme activities
(no-till with residue retention) (CAM) versus traditional agricultural management (moldboard plowing
Soil nitrogen and phosphorus without residue retention) (TAM) to a Haplic Cambisol soil in the North China Plain (NCP). Our results
Soil organic carbon showed that CAM significantly enlarged the stocks of SOC, DOC, LOC, STN and SAP in the 0–10 cm layer,
increased the contents of SOC, STN and SAP in the sand fractions, and promoted all of the enzyme activ-
ities in the bulk soil and all of the four particle-size fractions. Our results suggested that CAM increased
the nutrient contents in the sand fractions by both enlarging the content of particulate organic matter
and enhancing the activities of enzymes involved in nutrient cycling in these fractions. On the contrary,
the contents of SOC and nutrients in the silt and clay fractions were relatively resistant to the conversion
from CAM to TAM, indicating the limitation of CAM for stable SOC sequestration.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction
Traditional agricultural management (moldboard plowing
without residue retention) (TAM) is dominant in the North China
Plain (NCP), an intensive agriculture area with 320,000 km2 and
over 200 million population. This agricultural strategy has resulted
in serious environmental problems, such as groundwater contami-
nation (Hu et al., 2006) and soil organic carbon (SOC) loss in the NCP
(Wang et al., 2007). Conservational agricultural management (no-
till with residue retention) (CAM) has recently been expanded in
the NCP to increase crop yields whilst reducing soil erosion and
environmental degradation (Wang et al., 2007), compared with
TAM.
Tillage and residue management play an important role in SOC
turnover, aggregate formation and microbial abundance in soil
(Lagomarsino et al., 2009). No-till and crop residue retention tend
to decrease the turnover rate of macro-aggregate (Six et al., 2000),
∗ Corresponding author. Tel.: +86 311 85814360, fax: +86 311 88150953.
E-mail address: meimei841114@163.com (C. Hu).
resulting in SOC sequestration in the surface soil under CAM (Six et
al., 2002; Pacala and Socolow, 2004).
Soil enzymes produced by microorganisms play a key role in
the process of SOC mineralization (Ahn et al., 2009). They catalyze
the decomposition of different organic substrates, release plant
nutrients and influence whether SOC is depleted or sequestrated
(Fansler et al., 2005). Enzyme activities are unequally distributed
among soil particle-size fractions (Poll et al., 2003; Saviozzi et
al., 2007). In general, the carbohydrases [␤-glucosidase (␤-G),
␤-cellobiohydrolase, N-acetyl-␤-glucosaminidase (N-G) and ␤-
xylosidase] are reduced from the coarse sand to the clay fraction
(Stemmer et al., 1998; Kandeler et al., 1999a; Marx et al., 2005),
while the enzymes involved in nitrogen (urease and protease),
in phosphorus (acid and alkaline phosphatase) and in sulphur
(arylsulfatase) transformations, and the microbial biomass are
predominant in both the silt and clay fractions (Kandeler et al.,
1999b; Marx et al., 2005; Saviozzi et al., 2007). Similar to the
enzyme activities, SOC is also heterogeneously distributed among
particle-size fractions (Wu et al., 2006; Lagomarsino et al., 2009).
Mineral sorption is important to protect SOC from microbial attack
(Allison and Jastrow, 2006). The quartz particles that are domi-

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Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
conservational versus traditional agricultural management. Appl. Soil Ecol. (2010), doi:10.1016/j.apsoil.2010.03.007
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nant in the sand fractions have only weak bonding affinities to
SOC, while the clay particles, such as sesquioxides and layer sil-
icates, provide a large surface area and numerous reactive sites
to adsorb SOC through strong ligand exchange and polyvalent
cation bridges (Sposito et al., 1999). Consequently, SOC within
the sand, silt and clay fractions are allocated into the active,
intermediate and passive pool, respectively (Von Lützow et al.,
2007). These organo-mineral pools have different microbial avail-
ability (Marx et al., 2005) and can be separated by the physical
fractionations (Stemmer et al., 1998), such as wet-sieving and cen-
trifugation.
The distributions of SOC and microbial activities in particle-size
fractions are important for determining how agricultural practices
influence soil nutrient release and SOC balance (Kandeler et al.,
1999a; Saviozzi et al., 2007; Lagomarsino et al., 2009). A limited
number of papers investigated the distributions of SOC and micro-
bial activities in soil particle-size fractions under different tillage
or/and fertilization management (Kandeler et al., 1999b; Poll et al.,
2003; Saviozzi et al., 2007; Lagomarsino et al., 2009). However, few
of them linked the micro-scale enzyme activities to soil nutrient
contents and SOC balance in response to both tillage and residue
managements.
By investigating the soil enzyme activities involved in C, N, and
P transformations [␤-G, N-G, protease, urease and alkaline phos-
phomonoesterase (APH)], and the major nutrient contents [SOC,
total N (STN) and available P (SAP)] in the particle-size fractions of
a Haplic Cambisol in the NCP, our objective was to test whether the
different tillage and residue management modified the microbial
activities associated with the nutrient release and SOC balance at
the primary particle scale.
2. Materials and methods
2.1. Description of field site
This study was conducted on an 8-year conservational agri-
culture experimental field (37.90◦ N, 114.67◦ E, elevation 50 m),
which is located at the piedmont of the Taihang Mountains, in
the NCP and belongs to the Luancheng Agro-ecosystem Station,
the Chinese Academy of Sciences. The annual mean solar radia-
tion, temperature, precipitation and frost-free period in the station
is 547 kJ cm−2 , 12 ◦ C, 539 mm and 200 days, respectively. The soil is
classified as a silt loam Haplic Cambisol (IUSS Working Group WRB,
2006). In this NCP region, the prevalent cropping system is winter
wheat (middle October to early June) with summer corn (early June
to late September).
2.2. Experiment design and soil sampling
Two contrary agricultural management systems were estab-
lished in this study since 2001: (1) CAM [chopping and spreading
maize stalk (mean 11.26 Mg ha−1 year−1 ) after harvest, following
by wheat seeding with no-till seeders]; (2) TAM (removing out
maize stalk from field after harvest, following by 20 cm mold-
board plowing and wheat seeding with traditional seeders). Each
management treatment had three replicated plots. The size of
each plot was 30.0 m × 45.0 m. Fertilizer and irrigation were iden-
tical for both management systems. Before winter wheat sowing,
177 kg ha−1 urea and 326 kg ha−1 diammonium hydrogen phos-
phate were applied. Shortly after jointing, both wheat and corn
were fertilized at the rate of 193 kg urea ha−1 by surface broad-
cast. All plots were irrigated at sowing. Depending on rainfall,
additional three or four irrigations for wheat and two or three
irrigations for corn were applied using a sprinkler system. The
irrigation schedule was determined by soil moisture. Generally
Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
conservational versus traditional agricultural management. Appl. Soil Ecol. (2010), doi:10.1016/j.apsoil.2010.03.007
40–50 mm was applied for each irrigation when the soil moisture
of root zone declined to 60–65% of the field water-holding capacity
(FWC).
Soil samples for SOC, STN, SAP and microbial activity assays
were collected on October 6, 2008. Additionally, another soil sam-
pling was conducted on February 20, 2010 for soil basal respiration
(SBR), dissolved organic C (DOC) and light fraction organic C (LOC)
measurements. Five surface soil cores (0–10 cm) were collected
randomly from each plot after removing surface litters. This sam-
pling depth was used for the reason that the tillage effects on soil
microbial activities tend to be more significant near the surface than
those in the deeper layers (Muruganandam et al., 2009). The five
soil cores were mixed into one sample, stored in an ice box and then
in a cold room. The soil samples were ground through a 2-mm sieve
and divided into two sub-samples. One sub-sample was stored at
field moisture (75% of FWC) at 4 ◦ C for chemical and biochemical
analyses and the other was dried at 105 ◦ C for gravimetrical water
content.
2.3. Soil particle-size fractionation
Soil particle-size fractions were obtained by a combination of
wet-sieving and centrifugation after dispersion by the low-energy
sonication (Stemmer et al., 1998). This fractionation procedure is
less destructive to SOC and enzyme activities than the chemical
procedures (Stemmer et al., 1998). Field moist soil samples (35 g
equivalent dry weight) were dispersed in 100 ml of cooled dis-
tilled water (4 ◦ C) by a probe-type ultrasonic disaggregator (50 W,
120 s, probed at 15 mm depth). The coarse sand (2000–200 ␮m) and
fine sand (200–63 ␮m) fractions were recovered by wet-sieving.
The silt-size particles (63–2 ␮m) were separated by centrifuging
the remaining suspension at 150 × g for 2 min, and the pellet was
re-suspended and centrifuged again under the same condition.
This procedure was repeated twice to purify the silt fraction. The
remaining supernatants were centrifuged at 3900 × g for 30 min
to separate the clay-size particles (2–0.1 ␮m). The separated frac-
tions were divided into two sub-samples. One was dried at 105 ◦ C
for 24 h to calculate the recovery of fractionation. The other was
freeze-dried and stored at 4 ◦ C for chemical and biochemical anal-
yses within 4 weeks. In order to test the validity of this fractionation
method, a standard sedimentation method (Rowell, 1994) was con-
ducted on the same soil.
2.4. Soil chemical and biochemical assays
SOC (mg g−1 dry soil) was determined by the dichromate diges-
tion method (Kalembassa and Jenkinson, 1973), STN (mg g−1 dry
soil) by the semi-microkjedahl method (Nelson and Sommers,
1980), MBN (␮g g−1 dry soil) by Kandeler et al. (1999b), and SAP
(␮g P g−1 dry soil) by Olsen et al. (1954).
SBR (mg CO2 -C kg−1 dry soil d−1 ) was determined by the method
of Cookson et al. (2008). The majority (more than 80%) of DOC and
LOC were lost during the wet-sieving procedure. Consequently, we
determined the bulk stocks of DOC (kg ha−1 ) and LOC (Mg ha−1 ) in
the 0–10 cm layer using the method of Cookson et al. (2008).
The activities of APH (␮mol p-nitrophenol h−1 g−1 dry soil), N-
G (␮mol p-nitrophenol h−1 g−1 dry soil), ␤-G (␮mol p-nitrophenol
h−1 g−1 dry soil), protease (␮g tyrosine h−1 g−1 dry soil) and urease
(␮g NH4 + h−1 g−1 dry soil) were determined. The methods used to
determine these biochemical parameters were shown in Table 1.
All enzyme activities were determined in triplicate. The percentage
contribution of each parameter in each particle-size fraction to the
bulk soil was calculated based on the method of Stemmer et al.
(1998). The ratios of microbial parameters to MBN were calculated
based on the method of Landi et al. (2000).
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Table 1
The methods used for soil enzyme activity assays.

Enzyme Substrate (concentration) Optimum PH Reference

Alkaline phosphomonoesterases p-Nitrophenyl phosphate (10 mM) 10.0 Eivazi and Tabatabai (1977)
N-acetyl-␤-glucosaminidase p-Nitrophenyl-N-acetyl-␤-d-glucosaminidemine (10 mM) 5.5 Parham and Deng (2000)
␤-Glucosidase p-Nitrophenyl-␤-d-glucopyranoside (10 mM) 6.0 Tabatabai (1994)
Protease Casein (1%) 9.0 Ladd and Butler (1972)
Urease Urea (1 M) 8.0 Nannipieri et al. (1980)

2.5. Statistical analysis
Data (means ± SE) were compared by the LSD test at P < 0.05.
All statistical analyses were conducted with the software SPSS 13.0
(SPSS, 2004).
3. Results
3.1. Particle-size fractionation
The sonication method and the classical sedimentation method
yielded similar particle-size distributions (Table 2). The proportions
of particle-size fractions were not significantly different between
CAM and TAM (Table 2). The recovery rate of the sonication frac-
tionation procedure exceeded 99% (Table 2).
3.2. SOC, STN, SAP and C/N ratio in bulk soil and particle-size
fractions
The contents of SOC and SAP in the bulk soil were significantly
greater under CAM than those under TAM (Fig. 1). On the contrary,
STN content in the bulk soil was not significantly affected by differ-
ent managements (Fig. 1). The contents of SOC, STN and SAP in both
the coarse sand (2000–200 ␮m) and the fine sand (200–63 ␮m)
fractions were also significantly greater under CAM than those
under TAM, whilst no significant difference in those parameters in
either the silt (63–2 ␮m) or the clay (2–0.1 ␮m) fraction between
the two management treatments (Fig. 1). Under the same man-
agement, the contents of SOC, STN, and SAP increased with the
reduction of particle size (Fig. 1). The Recovery rates of SOC, STN
and SAP exceeded 99% (Table 3). The C/N ratio was similar in the
bulk soil, in the silt and the clay fractions between the two man-
agement treatments, but it was significantly greater in the coarse
and fine sand fractions under CAM than that under TAM (Table 2).
Under the same management, the C/N ratio significantly decreased
with the reduction of particle size (Table 2).
3.3. Soil microbial properties in bulk soil and particle-size
fractions
In the bulk soil, MBN and SBR, and the activities of APH, N-G, ␤-
G, protease and urease were significantly higher under CAM than
those under TAM (Figs. 1 and 2). These parameters were also sig-
nificantly higher in all of the four soil particle-size fractions under
CAM than those under TAM, except for MBN in the clay fraction and
SBR in the coarse sand, clay and silt fractions (Figs. 1 and 2). Under
the same management, MBN and SBR, and the activities of APH,
protease, and urease tended to increase, while the activities of N-G
and ␤-G decreased with the reduction of particle size (Figs. 1 and 2).
The average recovery rates of APH, SBR, MBN, N-G, ␤-G, protease
and urease were 105.0%, 98.0%, 99.6%, 72.9%, 88.1%, 104.6% and
103.2%, respectively (Table 3). The correlation coefficients between
SOC and microbial parameters were more remarkable in coarse and
fine sand fractions than those in silt and clay fractions (Table 4).
The activities of APH, ␤-G, N-G, urease and protease were signifi-
cantly correlated with each other in all of the particle-size fractions
(Table 4). CAM significantly promoted the ratios of SBR and enzyme
activities to MBN in the sand fractions, but not in the bulk soil and
the silt and clay fractions (Table 5).

3.4. Soil nutrients stock

The stocks of SOC, STN, SAP, DOC and LOC in the surface soil
(0–10 cm) were significantly greater under CAM than those under
TAM (Table 6).

4. Discussion

Compared with the chemical fractionation procedure (chemical

Fig. 1. Contents of soil organic C (SOC), available P (SAP), total N (STN) and basal
respiration (SBR) in the bulk soil and the particle-size fractions under conservational
(CAM) and traditional agricultural management (TAM). Data are means ± SE (n = 3)
and different letters above bars indicate significant difference (P < 0.05) between
managements (x, y) for each particle-size fraction and among particle-size fractions
(a, b, c, d, e) for each treatment, respectively.
dispersion + sedimentation), the ultrasonic procedure (ultrasonic
dispersion + centrifugation) has less effect on soil biochemical
properties (Stemmer et al., 1998). However, this ultrasonic pro-
cedure is not able to completely disperse soil aggregates (Marx et
al., 2005; Von Lützow et al., 2007). This was proven by the result
that the percentages of the sand fractions obtained by the ultra-
sonic procedure were slightly higher than those obtained by the
chemical procedure (Table 2). The high recovery rates of SOC, STN
and SAP indicated the validation of the method of Stemmer et al.
(1998). The recovery rates of ␤-G (88.1%) and N-G (72.9%) were rel-

Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
conservational versus traditional agricultural management. Appl. Soil Ecol. (2010), doi:10.1016/j.apsoil.2010.03.007
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Table 2
Soil particle-size distributions (percent of bulk soil) determined by the ultrasonic method (ultrasonic dispersion + centrifugation) and the standard chemical method (chemical
dispersion + Sedimentation) and C/N ratio (soil organic C/total N) under the conservational (CAM) versus traditional agricultural management (TAM).

Managements Fractionation method Particle size fractions Recovery (%)

Coarse sand Fine sand Silt Clay

Particle size distribution

CAM Ultrasonic dispersion + centrifugation 3.8 ± 1.2a, ␦ 10.3 ± 1.5a, ␥ 71.9 ± 1.0a, ␣ 13.9 ± 1.1a, ␤ 99.9
Chemical dispersion + Sedimentation 3.0 ± 1.0a, ␦ 9.6 ± 1.2a, ␥ 74.2 ± 2.0a, ␣ 13.2 ± 0.2a, ␤ 100.0

TAM Ultrasonic dispersion + centrifugation 3.2 ± 1.0a, ␦ 10.9 ± 0.7a, ␥ 72.2 ± 1.0a, ␣ 13.6 ± 0.9a, ␤ 98.9
Chemical dispersion + Sedimentation 3.8 ± 1.5a, ␦ 9.1 ± 0.7a, ␥ 72.5 ± 1.0a, ␣ 13.6 ± 0.9a, ␤ 97.0

Managements Fractionation method Particle size fractions In bulk soil

Coarse sand Fine sand Silt Clay

C/N ratio in particle size fractions

CAM Ultrasonic dispersion + centrifugation 15.0 ± 1.1a, ␣ 16.3 ± 1.4a, ␣ 9.5 ± 0.9a, ␤ 5.3 ± 1.0a, ␦ 8.0 ± 0.9a, ␥
TAM Ultrasonic dispersion + centrifugation 12.2 ± 1.9b, ␤ 14.7 ± 2.6b, ␣ 10.1 ± 1.3a, ␤ 5.3 ± 0.7a, ␦ 8.2 ± 1.1a, ␥

Data (means ± SE, n = 3) followed by the different letters in columns (a, b) and in rows (␣, ␤, ␥, ␦) indicate significant difference (P < 0.05).

atively low (Table 3), probably due to the fact that ␤-G and N-G were
predominately associated with coarse and fine sand fractions and
therefore poor protected during the fractionation procedure. On the
contrary, the recovery rates of APH, urease and protease exceeded
100% (Table 3), probably due to the liberation and activation of
enzymes from soil aggregates by sonication and wet-sieving.
4.1. Management effects on carbon balance and relevant enzyme
activities in soil particle-size fractions
SOC balance is mainly determined by the net result of car-
bon input and decomposition rate in soil (Silveira, 2005). Carbon
input under CAM tends to increase due to crop residue retention,
while the mineralization rate is more a function of the environ-
ment (Koarashi et al., 2009). The increase in bulk SOC content under
CAM (Fig. 1) was consistent with Li et al. (2007) who reported that
the SOC content in the surface layer (0–10 cm) was significantly
increased under 15-year CAM in the northern China. The increase
of SOC under CAM was probably resulted from the residue returned
into soil (Arshad et al., 1990). This was true for our study that
11.26 Mg ha−1 year−1 aboveground crop residue had been returned
under CAM. The 0–10 cm stocks of DOC and LOC were significantly
higher under CAM than those under TAM (Table 6). This result
agreed with Cookson et al. (2008) who reported that residue reten-
tion increased the contents of DOC and LOC in the surface soil
(0–10 cm) in Western Australia. These results indicated that CAM
had the potential to increase labile SOC sequestration. The quantity
and quality of organic matter returned into soil might determine its
distribution in different particle-size fractions (Lagomarsino et al.,
2009). The crop residues generally have a high availability to soil
microorganisms and consequently can be easily incorporated into
macro-aggregate (2000–250 ␮m) fractions during their decompo-

Table 3
The percentage contribution (%) of the parameters in each particle-size fraction to the bulk soil and the total recovery rates of parameters under CAM and TAM.

Parameter CAM TAM Particle-size fraction Parameter CAM TAM

Mean SE Mean SE Mean SE Mean SE

SOC 2.1 0.4 1.0 0.1 2000–200 ␮m APH 2.2 0.2 1.79 0.3
6.4 0.2 6.6 0.2 200–63 ␮m 6.4 0.4 7.98 0.5
68.3 3.8 69.9 6.8 63–2 ␮m 83.1 5.3 71.4 9.6
22.3 1.4 22.2 2.8 2–0.1 ␮m 19.1 1.2 18.1 1.4
99.1 99.8 Total 110.8 99.3

STN 1.1 0.3 0.7 0.1 2000–200 ␮m ␤-G 5.48 0.4 5.5 0.5
3.3 0.8 2.6 0.3 200–63 ␮m 16.9 1.1 10.5 0.9
60.5 5.4 60.4 3.1 63–2 ␮m 53.9 2.6 57.2 8.0
35.0 3.4 36.2 4.2 2–0.1 ␮m 13.3 1.2 13.2 0.9
99.9 99.9 Total 89.6 86.5

SAP 0.7 0.1 0.35 0.1 2000–200 ␮m N-G 4.0 0.2 3.7 0.2
3.9 0.3 2.3 0.3 200–63 ␮m 12.4 0.7 11.3 1.5
73.8 4.7 75.0 8.1 63–2 ␮m 43.3 1.5 50.8 3.9
22.4 1.5 22.2 1.3 2–0.1 ␮m 9.2 0.4 10.8 0.8
100.9 99.9 Total 69.1 76.7

MBN 0.85 0.3 0.5 0.0 2000–200 ␮m Protease 3.61 0.1 2.5 0.3
2.8 0.7 2.5 0.3 200–63 ␮m 4.51 0.3 5.5 0.5
77.7 3.4 72.9 5.1 63–2 ␮m 72.5 5.8 77.1 8.0
18.2 2.0 23.5 2.0 2–0.1 ␮m 23.3 3.3 20.1 1.8
99.7 99.5 Total 104.0 105.2

SBR 2.2 0.2 2.1 0.3 2000–200 ␮m Urease 2.2 0.1 2.1 0.2
8.1 0.6 7.8 0.4 200–63 ␮m 8.23 0.5 10.1 0.7
69.1 5.1 72.1 6.3 63–2 ␮m 79.0 4.9 74.7 6.6
16.6 1.3 17.8 0.8 2–0.1 ␮m 15.6 1.5 14.2 1.1
96.1 99.9 Total 105.1 101.2

Abbreviations: SOC, soil organic C; STN, soil total N; SAP, soil available P; MBN, microbial biomass N; SBR, soil basal respiration; APH, alkaline phosphomonoesterase; ␤-G,
␤-glucosidase; N-G, N-acetyl-␤-glucosaminidase.

Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
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Table 4
Correlation coefficients between properties in the particle-size fractions.

SOC STN SAP MBN APH ␤-G N-G Protease Urease

In the 2000–200 m fraction

SOC 1 0.72 0.81 0.81 0.75 0.83 0.86 0.87 0.86
STN 1 0.70 0.67 0.75 0.83 0.67 0.81 0.85
SAP 1 0.91 0.90 0.91 0.90 0.96 0.87
MBN 1 0.85 0.83 0.92 0.91 0.87
APH 1 0.88 0.86 0.76 0.84
␤-G 1 0.84 0.87 0.79
N-G 1 0.94 0.91
Protease 1 0.84
Urease 1

In the 200–63 m fraction

SOC 1 0.75 0.82 0.66 0.77 0.82 0.83 0.37 0.76
STN 1 0.79 0.78 0.76 0.81 0.92 0.80 0.73
SAP 1 0.89 0.79 0.75 0.89 0.49 0.71
MBN 1 0.84 0.86 0.93 0.82 0.94
APH 1 0.77 0.76 0.82 0.88
␤-G 1 0.79 0.75 0.84
N-G 1 0.97 0.76
Protease 1 0.73
Urease 1

In the 63–2 m fraction

SOC 1 0.69 0.76 0.73 0.76 0.76 0.78 0.85 0.33
STN 1 0.76 0.57 0.29 0.28 0.21 0.54 0.75
SAP 1 0.29 0.21 0.33 0.10 0.26 0.70
MBN 1 0.59 0.67 0.43 0.57 0.68
APH 1 0.82 0.76 0.74 0.87
␤-G 1 0.75 0.69 0.76
N-G 1 0.73 0.84
Protease 1 0.78
Urease 1

In the 2–0.1 m fraction

SOC 1 0.68 0.75 0.86 0.26 0.46 0.44 0.59 0.79
STN 1 0.76 0.91 0.56 0.78 0.65 0.75 0.89
SAP 1 0.74 0.80 0.63 0.74 0.76 0.57
MBN 1 0.54 0.67 0.49 0.71 0.49
APH 1 0.85 0.79 0.91 0.76
␤-G 1 0.88 0.94 0.87
N-G 1 0.75 0.93
Protease 1 0.76
Urease 1

For abbreviations, see Table 3. Numbers in bold are significant (P < 0.05) correlations, n = 9.

Table 5
Ratios of soil basal respiration (SBR) and enzyme activities to microbial biomass N (MBN) in bulk soil and particle-size fractions under conservational (CAM) and traditional
agricultural management (TAM).

Ratios Bulk 2000–200 ␮m 200–63 ␮m 63–2 ␮m 2–0.1 ␮m

CAM TAM CAM TAM CAM TAM CAM TAM CAM TAM

SBR:MBN (g CO2 -C g−1 MBN d−1 ) 0.5x, b 0.5 x, c 1.2 y, a 2.7 x, a 1.3 y, a 1.78 x, b 0.43 x, b 0.6 x, c 0.4 x, b 0.4 x, c
APH:MBN (␮mol PNP g−1 MBN h−1 ) 7.4 x, c 7.0 x, c 15.9 y, a 27.2 x, a 13.6 y, b 16.6 x, b 6.6 x, c 5.96 x, c 6.4 x, c 4.5 x, d
␤-G:MBN (␮mol PNP g−1 MBN h−1 ) 11.2 x, b 11.4 x, c 59.32 y, a 102.4 x, a 54.3 y, a 33.96 x, b 6.46 x, c 6.56 x, d 6.7 x, c 4.7 x, d
N-G:MBN (␮mol PNP g−1 MBN h−1 ) 11.2 x, c 10.6 x, c 44.6 y, a 64.0 x, a 39.73 x, b 33.96 x, b 5.2 x, d 5.46 x, d 4.7 x, d 3.6 x, e
Protease:MBN (␮g tyrosine g−1 MBN h−1 ) 20.7 y, b 26.9 x, c 74.3 y, a 108.0 x, a 26.9 y, b 42.3 x, b 15.9 x, d 20.8 x, d 21.8 x, c 16.8 x, e
Urease:MBN (g NH4 + g−1 MBN h−1 ) 0.6 x, b 0.8 x, c 1.3 y, a 3.3 x, a 1.4 y, a 2.2 x, b 0.5 x, b 0.6 x, c 0.4 x, b 0.4 x, d

For abbreviations, see Table 3. Data (means ± SE, n = 3) followed by the different letters indicate significant difference between CAM and TAM (x, y) for each fraction and
among different particle-size fractions (a, b, c, d and e) for each management, respectively.

Table 6
Bulk density and C, N and P stocks in the surface soil (0–10 cm) under the conservational (CAM) and traditional agricultural management (TAM).

Managements Bulk density (Mg m−3 ) SOC (Mg ha−1 ) DOC (kg ha−1 ) LOC (Mg ha−1 ) STN (Mg ha−1 ) SAP (kg ha−1 )

CAM 1.45 ± 0.02 a 15.14 ± 1.1 a 76.82 ± 8.4 a 0.45 ± 0.12 a 1.89 ± 0.11 a 61.3 ± 3.8 a
TAM 1.34 ± 0.05 b 13.93 ± 1.6 b 54.52 ± 2.1 b 0.33 ± 0.05 b 1.53 ± 0.08 b 49.6 ± 5.2 b

Abbreviations: SOC, soil organic C; DOC, dissolved organic C; LOC, light fraction organic C; STN, soil total N; SAP, soil available P. Data (means ± SE, n = 3) followed by the
different letters in columns (a, b) are significantly different (P < 0.05).

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Fig. 2. Activities of alkaline phosphomonoesterase, N-acetyl-␤-glucosaminidase, ␤-glucosidase, protease, urease, and microbial biomass N content in the bulk soil and the
particle-size fractions under conservational (CAM) versus traditional agricultural management (TAM). Data are means ± SE (n = 3) and different letters above bars indicate
significant difference (P < 0.05) between management treatments (x, y) for each particle-size fractions and among particle-size fraction (a, b, c, d, e) for each management
treatment, respectively.

sition (Stemmer et al., 1998), contributing to the high SOC contents
and C/N ratios in the coarse and fine sand fractions under CAM
(Table 2). More than 68% and 22% of the total SOC were existed
in the silt and clay-size fraction, respectively (Table 3). However,
the SOC contents in these fractions were not significantly differ-
ent between CAM and TAM (Fig. 1). These results indicated the
resistance of SOC in these fractions to management modifications.
The silt and clay fractions had rather low C/N ratios (average 7)
(Table 2). This suggested the high degree of SOC humification in
these fractions (Stemmer et al., 1998).
Besides the carbon input, the SOC decomposition rate is another
important factor influencing SOC balance. SBR directly indicates the
SOC decomposition rate. Some enzymes involved in soil C cycling,
such as ␤-G and N-G, can provide the information on SOC decom-
position potential (Allison and Jastrow, 2006). The ␤-G catalyzes
the final step of cellulose degradation and N-G hydrolyzes the glu-
cosamine (Reboreda and Caçador, 2008). The activities of both ␤-G
and N-G tended to decrease (Fig. 2), while the MBN tended to
increased, from the coarse sand to the clay fraction (Fig. 1). These
trends agreed with Marx et al. (2005) who explained that the large
amount of polymeric material in the sand fractions could account
for the high carbohydrase activities in these fractions. The higher
content of MBN in the finer fractions was probably due to the
microorganism protection from faunal predation (Poll et al., 2003).
CAM significantly increased SBR in the bulk soil and the fine sand
fraction, and increased MBN and the activities of N-G and ␤-G in all
of the four particle-size fractions, especially in the sand fractions
(Figs. 1 and 2). These results indicated that the accumulated SOC in
the sand fractions under CAM had a high decomposition potential.
To sum up, our results showed that CAM increased the SOC stock
in the surface soil layer (0–10 cm) (Table 6). However, the accu-
mulated SOC was mainly derived from the coarse and fine sand
fractions rather than from the silt and clay fractions (Fig. 1), suggest-
ing the limitation of CAM for SOC sequestration. Furthermore, the
C/N ratios and enzyme activities in the sand fractions were higher
under CAM than those under TAM. These results indicate that the
accumulated C under CAM was easy to be decomposed, which fur-
ther limited the potential of CAM for stable SOC sequestration.
4.2. Management effects on the nutrient contents and relevant
enzyme activities in soil particle-size fractions
SOC is the major source of plant nutrients (Bauer and Black,
1994). Its quantity and quality directly influence the availability
of soil nutrients, such as nitrogen and phosphorus. The increases
of STN and SAP in the sand fractions (2000–200 and 200–63 ␮m)
under CAM (Fig. 1) were probably resulted from the residue reten-
tion (Arshad et al., 1990). The increasing trend of STN from coarse
sand to clay fractions (Fig. 1) agreed with Marx et al. (2005). The
SAP determined by the Olsen method is a valid parameter for esti-
mating the P availability in soil (Demetz and Insam, 1999). The
SAP content tended to increase and be less susceptible to man-
agement modifications from coarse sand to clay fraction (Fig. 1).
These results suggested that CAM increased P availability in the
sand fractions, which could increase the crop yields. On the con-
trary, phosphorus in the silt and clay fractions was associated with
mineral particles and hardly modified by management (Fig. 1).
This kind of phosphorus had poor availability to plants and soil
microorganisms. The processes of nutrient release from SOC are
regulated by soil enzyme activities (Dorodnikov et al., 2009). There-
fore, soil enzymes involved in nutrient cycling have been suggested
as potential indicators of soil quality (Cardelli et al., 2004). Ure-
ase and protease play important roles in organic N mineralization
since they hydrolyze urea into ammonium and hydrolyze protein
into amino acid, respectively. Phosphomonoesterase is important
to degrade organic phosphorus. Acid phosphomonoesterase dom-
inates in acid soils, while APH dominates in alkaline soils (Eivazi
and Tabatabai, 1977). In the present study, the soil is weak alka-
line (pH 7.6). So we determined the APH activities in particle-size
fractions. In the sand fractions, activities of urease, protease and
APH were significantly higher under CAM than those under TAM
(Fig. 2), indicating the acceleration of nutrient cycling in these frac-

Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
conservational versus traditional agricultural management. Appl. Soil Ecol. (2010), doi:10.1016/j.apsoil.2010.03.007
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S. Qin et al. / Applied Soil Ecology xxx (2010) xxx–xxx
tions. This accounted for the larger stocks of SAP and STN in the
surface layer (0–10 cm) of soil under CAM, compared with those
under TAM (Table 6). On the contrary, the increase of these enzyme
activities in the silt and clay fractions did not significantly increase
the nutrient contents in these fractions (Fig. 1), probably due to the
fact that some minerals, such as allophane, may sorb and stabilize
soil enzymes (Allison, 2006). These stabilized enzymes are consid-
ered to have potential rather than in situ activities (Burns, 1982).
These results indicated that the enzyme activities in the silt and
clay fractions had little direct effects on nutrient cycling.
In a word, CAM promoted the microbial activities in all of
the four particle-size fractions. However, nutrient cycling was
increased only in sand fractions. It seemed that the increased
enzyme activities in the sand fractions contributed to the nutrient
release, while those in the silt and clay fractions did not.
4.3. Correlation between soil chemical and biochemical
properties in bulk soil and particle-size fractions
The Pearson correlation coefficients were analyzed between the
soil chemical and the microbial properties in different particle-
size fractions (Table 4). Significant liner correlations between
chemical and microbial parameters were widely reported in bulk
soil (Cookson et al., 2008). In the particle-size fractions, how-
ever, the correlation coefficients varied with both the fraction
and parameter. In general, significant correlations with all tested
microbial properties were observed for SOC in the coarse sand
(2000–200 ␮m) fraction, for STN in the coarse (2000–200 ␮m) and
fine (200–63 ␮m) sand fractions (Table 4). On the contrary, SOC
and STN were sometimes not significantly correlated with micro-
bial parameters in the silt and clay fractions (Table 4). These results
indicated that SOC in the sand fractions had high microbial avail-
ability, while that in the finer fractions were protected by mineral
particles and consequently had a low microbial availability. The
correlation coefficients between microbial parameters were sig-
nificant in all of the four fractions (Table 4). This was probably
attributed to the fact that extracellular enzymes accounted for con-
siderable part of enzyme activities in the silt and clay fractions.
Simmons and Coleman (2008) reported that soil nutrient concen-
trations were one of the factors that most heavily influenced the
abundance and diversity of soil microbes. Those significant corre-
lations in the sand fractions indicated the dependence of soil C, N
and P transformations on soil enzymes and the availability of sub-
strates for soil microbes to excrete such soil enzyme activities in
these fractions (Lagomarsino et al., 2009).
4.4. Ratios of enzyme activities to microbial biomass in bulk soil
and in the particle-size fractions
The ratios of SBR and enzyme activities to microbial biomass
(MBN) were significantly higher under CAM than those under TAM
in the coarse and fine sand fractions, though no significant differ-
ence existed in the bulk soil (Table 5). This result indicated that
the increases in enzyme activities under CAM were mostly orig-
inated from the enhancement of enzyme production rather than
the increase of microbial biomass. In general, the ratios decreased
with the decline of particle size (Table 5). This suggested that
microorganisms in the coarse and fine sand fractions were more
active in SOC decomposition than those in the silt and clay frac-
tions.
5. Conclusions
Enzyme production by microorganisms and stabilization by
physical processes influence SOC decomposition and nutrient
cycling (Martens, 2000). The separation of differently sized particle
Please cite this article in press as: Qin, S., et al., Soil organic carbon, nutrients and relevant enzyme activities in particle-size fractions under
conservational versus traditional agricultural management. Appl. Soil Ecol. (2010), doi:10.1016/j.apsoil.2010.03.007
7
fractions provided SOC and enzyme pools which differed signifi-
cantly in size and stability. Microorganisms within sand fractions
were active in SOC decomposition. Therefore, the accumulated par-
ticular organic carbon in the sand fractions under CAM significantly
increased the enzyme activities and consequently accelerated the
nutrient cycling in the sand fractions, resulting in the increase
of nutrient contents in these fractions. On the contrary, SOC and
enzymes within the silt and clay fractions were stabilized by min-
eral particles and consequently had limited capacity to release plant
nutrients. Although the enzyme activities in the silt and clay frac-
tions were significantly increased by CAM, this increase did not
bring greater contents of plant nutrients in these fractions. We
concluded that the conversion from TAM to CAM accumulated the
SOC content, prompted the microbial activities, and subsequently
increased the nutrient contents in the sand fractions. The 8-year
period of CAM was too short to induce modifications to the con-
tents of SOC and plant nutrients in the silt and clay fractions,
though the enzyme activities were significantly increased in these
fractions.
Acknowledgements
This work was supported by the National Natural Science Foun-
dation of China (30870456) and the Innovation Knowledge Project
of the Chinese Academy of Sciences (Grant Nos. KSCX2-YW-449
and KZCX2-YW-Q02-02). We are grateful to Xiaoqing Bi for soil
sampling.
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