R.C. Suelo Nutr. Veg.

10(2): 93 - 101 (2010)

SOIL GLYCOSIDASE ACTIVITIES AND WATER

SOLUBLE ORGANIC CARBON UNDER DIFFERENT
LAND USE TYPES
X. Z. Ma1,2, L. J. Chen*1, Z. H. Chen1, Z.J. Wu1, L.L. Zhang1, Y.L. Zhang1
1Institute
of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016 (China)
2Institute
of Soil Fertilizer and Environment Resource, Heilongjiang Academy of Agricultural
Sciences, Harbin 150086, China.
* Corresponding author: [email protected]
ABSTRACT
The purpose of this study was to measure the effects of different land uses on soil
glycosidase activities (α- and β-glucosidase, α- and β-galactosidase), water soluble
organic carbon (WSOC) and their relationships. Glycosidase activities showed
significant differences under different land use types, the highest one was woodland. β-
glucosidase had the highest activity among the four glycosidases. The activities of these
glycosidases decreased with increasing soil depth, being all significantly affected by
change of soil depth. Except grassland, the four glycosidase activities intercorrelated
each other. Woodland had the highest content of WSOC in the soil depth of 0-20 cm
and at increasing soil depth, WSOC content decreased sharply under woodland and
grassland. Glycosidase activities had positive and significant relationships with WSOC.
Glycosidase activities and WSOC all had significant correlations with soil total organic
carbon (TOC) and pH, which were sensitive to different land use types. We found that
glycosidase activity indirectly impacts on nutrient recycling and energy flow in soil
under different land use types.
Keywords: Glycosidase activities, Water soluble organic carbon, Land use, Soil depth.
INTRODUCTION
Different land use types not only had
effects on soil structure, soil erosion and
biodiversity (Crist et al., 2000), but also
on soil enzymes activities and soil
nutrient cyclings (Gewin et al., 1999;
Islam and Weil, 2000; Acosta-Martínez et
al., 2003). Enzymes catalyze all
biochemical reactions and are integral
part of nutrient cycling in soil. Soil
glycosidase is a group of hydrolases
involved in the hydrolysis of soil
glycosides, among which, α- and β-
glucosidase and α- and β-galactosidase
are the major members, widely distributed
in nature (Eivazi and Tabatabai, 1990)
and playing an important role in the
carbon cycle of soil ecosystem. β-
glucosidase was sensitive to different soil
management (Deng and Tabatabai, 1996;
Bandick and Dick, 1999). Many
researches had studied the effects of
tillage (Deng and Tabatabai, 1996;
Curci et al., 1997), crop rotation (Bandick
and Dick, 1999), fertilizer amendment
(Mijangos et al., 2006; Melero et al.,
2007; Sastre-Conde et al., 2007) on
glycosidase activities, but less studies are
present about the effects of different land
use types on glycosidase activities.
Among the components of soil carbon
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Soil glycosidase activities and WSOC under different land use types, Ma et al.
storage, water-soluble organic carbon
(WSOC) is the most dynamic C pool in
soils. It is only a small proportion of the
total organic matter in the soil, present in
soil solution and passing a filter pore size
of 0.45 μm (Herbert and Bertsch, 1995).
The WSOC could be used by microbes
quickly, and it is a useful indicator
reflecting the turnover rate of soil organic
matter. Many researches had studied its
response to fertilization and tillage
management (Mazzarino et al., 1993;
Erich and Trusty, 1997; Campbell et al.,
1999; Chantigny et al., 1999), but little is
know about its dynamics under different
land uses, and its relations with soil
glycosidase activities.
The present investigation has been
aimed at studying the effect of different
land use types on soil glycosidase
activities and WSOC. In addition,
relationships between glycosidase
activities and WSOC were also studied.
MATERIALS AND METHODS
Study site
Shenyang Experimental Station of
Ecology is a member of Chinese
Ecosystem Research Network (CERN)
under Chinese Academy of Sciences, and
locates in the Sujiatun District of
Shenyang City, Northeast China. This
station was established in 1990, with a
total area of about 15 hm2. Its soil is
classified as aquic brown soil.
The mean annual temperature is 7.0-
8.0ºC, mean annual precipitation is 650-
700 mm, and non-frost period is 147-164
days. Since its establishment, this station
installed four types of land use, i.e.,
lowland for rice, upland for corn,
grassland, and woodland for Populus
canadensis.
Soil sampling
Soil samples were taken at the depths of
0-5, 5-10, 10-20, 20-30, 30-40 and 40-50
cm from each type of the lands in March
2004 by using a core sampler of 5 cm in
diameter.
Four duplicates were installed for each
type of the lands, and each sample was a
composite of 5 cores. A portion of the
samples was air-dried for physical and
chemical analysis, and another portion
was kept fresh for the determination of
soil water soluble organic carbon content
and enzyme activities.
Soil total organic carbon (TOC) and
pH analysis
Soil pH was measured in soil: water
suspension (1:2.5 ratio) with glass
electrode (PSH-3C) (Lu, 2000); total
organic carbon was determined with Liqui
TOC analyzer (Elementar, German).
Determination of soil glycosidase
activity
Glucosidase and galactosidase activities
were determined as described by Eivazi
and Tabatabai (1988), glycosidase
activities were determined with pnitrophenyl-
glucopyranoside as substrate
(50 mmol L-1), with incubation at pH 6.0
(modified universal buffer, MUB) and
37ºC. After 1 h, 0.5 M CaCl2 and pH 12.0
MUB were added to precipitate humic
molecules responsible for brown
coloration and extract p-nitrophenol,
respectively.
The amount of p-nitrophenol released
by glycosidases was determined
colorimetrically at 410 nm (extinction
coefficient is 0.9998**). Glycosidase
activities were expressed as mg pnitrophenol
kg-1 soil h-1.
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R.C. Suelo Nutr. Veg. 10(2): 93 - 101 (2010)
Determination of soil water soluble
organic carbon (WSOC)
Soil water soluble organic carbon
(WSOC) was determined by shaking 50 g
field-moist soil with 150 mL deionized
water for 1 h (250 rpm), the suspension
was centrifuged at 10000 rpm for 10 min,
and the supernatant was collected with a
0.45 μm polycarbonate membrane filter
under vacuum (Chantigny et al., 1999).
The WSOC content was determined by
using Liqui TOC analyzer (Elementar,
German).
Statistical analysis
The results were analyzed statistically
adopting analysis of variance (ANOVA),
which were performed using SPSS 11.0
statistical package. Means separation was
using Fisher’s least significant difference
(LSD) test at p<0.05.
RESULTS
Total organic C (TOC) content and pH
value
TOC and pH value of the tested soils
under different land use types are shown
in Figure 1. There was the highest content
of soil organic matter under woodland,
followed by grassland, lowland and
upland, in the order listed. The content of
TOC decreased with soil depth increasing,
sharply in the upper layer (0-10 cm) under
woodland and grassland, then, changed
gently. The pH value was smaller in
upland than others, and was higher in the
whole soil profiles except 0-5cm under
grassland; it increased with soil depth
increasing under upland, lowland and
grassland, whereas, under woodland, it
decreased in 0-20 cm, then increased
below 20 cm.
0
10
20
30
40
50
4 5 6 7 8 9 10
pH
soil depth (cm)
lowland
upland
woodland
grassland
0
10
20
30
40
50
60
0 5 10 15 20 25 30 35
Total organic carbon (g kg-1)
soil depth (cm)
lowland
upland
woodland
grassland
Soil depth (cm)
Figure 1. Total organic carbon and pH value under different land use types.
Soil glycosidase activities
The activities of glycosidases were all
significantly affected by changes of soil
depth and land use and β-glucosidase had
the highest activity among them in the
soil profiles under different land use
types (Figure 2). Generally, they sharply
decreased by increasing the soil layer
from 0-5 cm to 5-20 cm, but gently in
deeper layers, displaying the same
distribution patterns observed for other
soil enzymes. Among the four types of
land use, woodland had the highest
95
Soil glycosidase activities and WSOC under different land use types, Ma et al.
0
10
20
30
40
50
60
0 10 20 30 40
α-galactosidase activity
(mg h-1 kg-1)
soil depth(cm)
0
10
20
30
40
50
60
0 20 40 60 80
0
10
20
30
40
50
60
0 5 10 15 20
α-glucosidase activity
(mg h-1 kg-1)
soil depth (cm)
10
20
30
40
50
60
0 50 100 150 200 250 300
β-glucosidase activity
(mg h-1 kg-1 )
upland
lowland
woodland
grassland
0
10
20
30
40
50
60
upland
lowland
woodland
grassland
20 40 60
β-galactosidase activity
(mg h-1 kg-1)
Soil depth (cm) Soil depth (cm)
0 50 100 150 200 250 300
Figure 2. Glycosidase activities in soil profile under different land use types.
activities of α- and β-glucosidase and β-
galactosidase in 0-10 cm soil layer,
followed by grassland, lowland and
upland, while lowland had a significantly
higher α-galactosidase activity in this
layer than other lands (Figure 2). The
relationships between glycosidase
activities and TOC, pH are shown in
Table 1.
There were positive and significant
relationships among glycosidases
activities and TOC content (p<0.001)
under four land use types, well, there
were negative and significant
relationships with soil pH (p<0.001) in
soil profiles except in woodland.
Meanwhile, except α-glucosidase, other
three glycosidases activities had positive
relationships with soil pH (p<0.05) in
woodland. The vertical distribution of
glycosidases activities had the close
relationships with soil characteristics
(TOC and pH) (Table 1). Linear
regression analysis of the activities of the
four enzymes showed that they were
significantly intercorrelated in woodland,
lowland and upland (Table 2). By
contrast, there was no significant
relationship between glucosidase and
galactosidase in grassland (Table 2).
Soil water soluble organic carbon
(WSOC)
Contents of soil water soluble organic
carbon (WSOC) decreased sharply
with soil depth increasing in woodland,
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R.C. Suelo Nutr. Veg. 10(2): 93 - 101 (2010)
Table 1. Correlation coefficients between soil glycosidase activities and soil total
organic carbon (TOC) and pH under different land use types.
Enzyme activities
Land use
types
Soil
characteristics
α-Glu β-Glu α-Gala β-Gala
TOC 0.794*** 0.899*** 0.843*** 0.817***
Lowland
pH -0.906*** -0.839*** -0.924*** -0.912***
TOC 0.759*** 0.778*** 0.758*** 0.756***
Upland
pH -0.825*** -0.737*** -0.783*** -0.882***
TOC 0.943*** 0.907*** 0.901*** 0.881***
Grassland
pH -0.535** -0.475* -0.462* n.s.
TOC 0.966*** 0.938*** 0.882*** 0.888***
Woodland
pH n.s. 0.464* 0.447* 0.470*
(Glu=glucosidase Gal=galactosidase) (* p<0.05, ** p<0.01, *** p<0.001, n.s. not significant)
grassland and lowland, but a smaller
fluctuation in upland (Figure 3).
Woodland had the highest WSOC content
in 0-20 cm, well, the lowland, grassland
and upland had lower content of WSOC.
Four land use types [except for upland
(n.s.)], there were significant and positive
correlations between glycosidase
activities and WSOC content (from
r=0.66*** to r=0.90***) (Table 3).
Results also showed that content of
WSOC under lowland, grassland and
woodland had significantly correlations
with TOC (p<0.01), but not significant in
upland; and there were negative and
significant correlations with pH under
lowland and grassland (p<0.01) (Table 4).
DISCUSSION
Total organic C (TOC) and pH value
Different land use types affected the input
and output of soil organic matter directly;
vegetation also had significant effect
on soil organic matter. Woodland always
had higher content of TOC than cropland,
due to many fallings back to soil, lots of
root distributed widely in soils of
woodland and grassland, so they had
much higher content of soil TOC in the
upper soil layer. Besides, the cultivation
accelerated the decomposition of soil
organic matter, made it decrease sharply
(Davidson, 1986). Different land use
types not only affect the content and
distribution of C in soil directly, also had
effects on some microbial conditions,
which had close relationships with
formation and transformation of C, then
affect nutrient distribution indirectly.
Soil glycosidase activities
Different land use types had different
levels of soil fertility. These changing
trends of glycosidases activities
decreasing with soil depth have been
reported in many researches (Eivazi and
Tabatabai, 1990; Deng and Tabatabai,
1996; Taylor et al., 2002), they were just
like the tendency of organic C in soil
profile, because these activities of
97
Soil glycosidase activities and WSOC under different land use types, Ma et al.
Table 2. Correlation coefficients (r)
between activities of paired enzymes in
soil samples under different land use
types.
Enzyme activities
Land use
types
β-Gala α-Glu β-Glu
α-Gala 0.994*** 0.881* 0.887*
Lowland β-Gala 0.886* 0.889*
α-Glu 0.998***
α-Gala 0.993*** 0.990*** 0.980**
Upland β-Gala 0.990*** 0.990***
α-Glu 0.961**
α-Gala 0.996*** n.s. n.s.
Grassland β-Gala n.s. n.s.
α-Glu 0.982***
α-Gala 0.989*** 0.988*** 0.967**
Woodland β-Gala 0.996*** 0.990***
α-Glu 0.988***
(*p<0.05, **p<0.01, ***p<0.001, n.s. not significant)
glycosidases in soil and content of
organic C always had significant and
positive relationships, which had been
proved by several investigators (Eivazi
and Tabatabai, 1988; Bandick and Dick,
1999; Marx et al., 2005), our study also
found the significant relationships
between activities of glycosidases and
TOC in the tested soils (Table 1).
Table 3. Relationships between soil
glycosidase activities and water soluble
organic carbon content.
Upland Lowland Grassland Woodland
α-glucosidase n.s. 0.68*** 0.84*** 0.77***
β-glucosidase n.s. 0.68*** 0.90*** 0.73***
α-galactosidase n.s. 0.66*** 0.81*** 0.73***
β-galactosidase n.s. 0.71*** 0.78*** 0.67***
(** p<0.01, *** p<0.001, n.s. not significant)
0
10
20
20 40 60 80 100 120
WSOC (mg C kg-1 soil)
soil depth(cm)
upland
lowland
woodland
grassland

Soil depth (cm)

WSOC
(mg C kg-1 soil)
20 40 60 80 100 120
0
30
40
50
60
woodland
grassland
upland
lowland
Figure 3. Water soluble organic carbon
content in soil profile under different land
use types.
Table 4. Correlation coefficients between
WSOC and soil properties (TOC, pH) in
soil profile under different land use types.
TOC pH
Lowland 0.633** -0.681***
Upland n.s. n.s.
Grassland 0.864*** -0.639**
Woodland 0.771*** n.s.
(** p<0.01, *** p<0.001, n.s. not significant)
In topsoil enzymes always had higher
activities than other soil depth, the main
reason maybe that there were higher
content of soil organic matter and
microbial biomass C, which would
stimulate the activity of microorganism,
and accelerate the rate of enzyme
synthesize (Ekenler and Tabatabai, 2003).
There were different enzyme activities
under different land use types, due to the
differences in organic C content among
soils (Acosta-Martínez et al., 2007).
The β-glucosidase activity had close
relationship with soil pH, which was
conformed to other studies (Eivazi and
Tabatabai, 1990; Wang and Lu, 2006),
they found that β-glucosidase activity
decreased with increasing pH from 4.3 to
7.4, 4.5 to 8.5 respectively, in this study it
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R.C. Suelo Nutr. Veg. 10(2): 93 - 101 (2010)
decreased with pH increasing from 5.4 to
7.8. However, Deng and Tabatabai (1996)
found the inconsistent relationship
between β-glucosidase activity and soil
pH, significant and positive correlation
between them. The main reason maybe
that soil pH influenced soil
microorganism, synthesis and secretion of
enzymes, also the stability of enzymes
(Wang and Lu, 2006). Besides, the
differences in enzyme activities found in
soil samples also may have been due to
the difference in soil pH, because the
rates of synthesis and release of these
enzymes by soil microorganisms are
related to soil pH (Deng and Tabatabai,
1996).
The α- and β-glucosidase and β-
galactosidase had higher activities under
woodland than other land use types. In
woodland, there were some kinds of litter
fallings remained on or in the soils,
higher content of organic matter, higher
enzymes activities. Investigators also had
found that enzymatic characteristic of soil
was very sensitive, and could do as a
potential quality index of soil system
(Bandick and Dick, 1999). Activity of β-
glucosidase was the highest one in this
study, which was conformed to former
studiers (Eivazi and Tabatabai, 1988;
Ekenler and Tabatabai, 2003), which
meant that β-glucosidase was sensitive to
changes of land use types. α- and β-
glucosidase and α-and β-galactosidase
activities were significantly
intercorrelated, suggest that glycosidase
have similar origin and persistence in soil
(Bandick and Dick, 1999; Acosta-
Martínez et al., 2007). We may conclude
that glucosidase and galactosidase have a
different origin under grassland.
Soil water soluble organic carbon
Soil water soluble organic carbon was
only fewer percents of soil total organic
carbon, but as the active part. Land uses
and management practices could affect
soil properties, and also influence WSOC.
Soil properties determine organic matter
solubility. WSOC content in soil profile
decreased with soil depth increasing,
maybe due to close relationships between
WSOC with soil total organic carbon,
which showed similar trends in soil
profiles. Generally, large numbers of
soluble organic matter were eluviated
from residues layer to mineral layer in
forest soil, so that amount of WSOC in
the topsoil was always higher than other
soil depths.
In general, WSOC concentrations
varied in different land uses, such as the
forest soils, grassland soils, arable soils
and so on, mostly due to different
vegetation types (Delprat et al., 1997;
Haynes, 2000). Besides, we also had
learned that fertilization could affect the
content of WSOC greatly (Zsolnay and
Görlitz, 1994; Jensen et al., 1997; Martin-
Olmedo and Rees, 1999). Chantigny
(2003) found that content of WSOC
decreased with increasing amount of
nitrogenous fertilizer as a result, upland
had higher content of WSOC than other
land uses. Another factor was moisture,
Christ and David (1996) found that the
total amount of WSOC leaching from
forest soil were increasing with times of
leaching, so that the woodland and
lowland had higher content of WSOC, the
other possible reason would be the
material returned to the soil by tree
canopy, which contained more lignin and
other recalcitrant compounds than
agricultural crop residues (Chantigny,
2003). The WSOC content in upland was
lower than others, due to the reasons of
fertilization and moisture (Christ and
David, 1996), the biochemical and
physical environment of upland were
different from other land uses. In this
study, there was significant and negative
correlation between WSOC and soil pH
in soil profile under lowland and
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Soil glycosidase activities and WSOC under different land use types, Ma et al.
grassland (p<0.01), while not in upland
and woodland.
CONCLUSIONES
The present results clearly showed that
different land use types had profound
impact on soil glycosidase activities and
content of water soluble organic carbon
(WSOC), which were all decreasing
with soil depth increasing. Besides, β-
glucosidase was most sensitive to
different land uses. Close relationships
among glycosidase activities, WSOC and
total organic carbon (TOC) contents,
meant that glycosidase activity would be
indicators for changing in soil quality,
which also indicated that effects of
different land uses on soil biological
activity are very important.
ACKNOWLEDGMENTS
This research was funded by Research
and Demonstration of Agricultural
Science (2007-3), Public Sector’s Special
Research of Ministry of Agriculture. The
authors thank Shenyang Experimental
Station of Ecology，which is a member
of Chinese Ecosystem Research Network
(CERN) under Chinese Academy of
Sciences, for its support to collect soil
samples.
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