Acta Ecologica Sinica 30 (2010) 123128

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Acta Ecologica Sinica

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

Nitrogen released from different plant residues of the Loess Plateau and their
additions on contents of microbial biomass carbon, nitrogen in soil
Zhou Jian-Bin *, Chen Xing-Li, Zhang Ying-Li, Liu Jian-Liang
College of Resource and Environment Science, Northwest A&F University, Yangling, Shaanxi 712700, China

a r t i c l e

i n f o

Keywords:
Loess Plateau
Plant residues
N mineralization
Soil microbial biomass carbon and nitrogen

a b s t r a c t
An incubation method was used to investigate the nitrogen release characteristics from the residue of ten
plant species which commonly grow in the northern part of the Loess Plateau. The effect of the residue on
soil microbial biomass carbon (SMBC) and soil microbial biomass nitrogen (SMBN) was also determined.
There were signicant differences in the total N content and the C/N ratios among the different types of
plant residue. The total N content of the residues ranged from 6.61 to 32.78 g kg 1. The C/N ratio of the
residue ranged from 14 to 65. There was an immediate increase in soil N after alfalfa, erect milkvetch, and
korshinsk peashrub residue was added to the soil. In contrast, soil N decreased after elm, sea buckthorn,
and wild peach residue was added to the soil. The soil N content remained relatively low for 1434 days
and then increased. This indicated that N immobilization occurred during the early portion of the incubation period when elm, sea buckthorn and wild peach residue was added to the soil. Soil N levels were
low during the entire incubation period when simon poplar, locust, Stipa bungeana, and old world bluestem residue were added to the soil. The addition of plant residue signicantly increased SMBC and SMBN
in all treatments. The SMBC and SMBN values were greatest in treatments containing plant residue with
high total N content and low C/N ratios. The C/N ratios of korshinsk peashrub, sea buckthorn, and wild
peach residues were similar, but the amount of N released from these residues and the effects of the residue on SMBC and SMBN in soil were signicantly different. This indicates that not only the C/N ratio but
also the chemical composition of the plant residue affected decomposition. It is important to consider C
and N release characteristics from plant residue in order to adjust the C and N balance of soil when revegetating degraded ecosystems.
2010 Ecological Society of China. Published by Elsevier B.V. All rights reserved.

1. Introduction
The amounts and types of plant residues returned into in terrestrial ecosystem affect the fertility of soil [1]. Microbes in ecosystem
play an important role in element and energy ows in soil ecosystem as the driving forces for transformation of plant residues in
soil; and nutrients in microbial biomass also involve in the nutrient
cycles in soil ecosystem [2]. Therefore, soil microbial biomass is
used as an important index to evaluate the effects of different cultivation and management on quality and function of soil ecosystem [36].
The Loess Plateau is a region suffering from very serious soil
erosion in the world due to the deteriorating vegetation by deforestation and inadequate cultivation management. Severe soil erosion results in the divided land, poor soil, and weak stabile
ecosystem. In recent years, ecological restoration is a key issue in
developing the west region of China [6]. Different measures of
reestablishing the vegetation in the region, such as change the
* Corresponding author.
E-mail address: jbzhou@nwsuaf.edu.cn (J.-B. Zhou).

marginal crop land into forest and grassland with trees, shrubs
and grass, are used to control soil and water erosion. There has a
series of researches to study the effects of ecological restoration
on soil physical and chemical properties in the region [7,8]. Nitrogen is a factor limited the plant growth in most ecosystems. However, the nitrogen release from the residues of these plants is less
understood, and there is also less research to study the effects of
addition of different residues on soil microbial biomass carbon
and nitrogen.
Therefore, the objectives of this study were to evaluate the effects of different residues of plant species from the Loess Plateau
on immobilization and mineralization of N in soil and on soil
microbial biomass carbon and nitrogen.

2. Materials and methods

2.1. Materials
The soil used was sampled from Shenmu Experimental Station
of Institute of Soil and Water Conservation, CAS, in fall of 2006.

1872-2032/$ - see front matter 2010 Ecological Society of China. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.chnaes.2010.04.001

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J.-B. Zhou et al. / Acta Ecologica Sinica 30 (2010) 123128

The previous crop was soybean (Glycine max (L.) Merr), and soil
was Typ-Los-Orthic Entisols. Soil was air-dried, and sieved in
2 mm. The basic physicalchemical properties of the soils were:
organic carbon, 4.87 g kg 1; total N, 0.57 g kg 1; NO3-N, 32.99
mg kg 1; NH4-N, 1.88 mg kg 1; pH 7.90; Olsen-P, 2.58 mg kg 1;
clay, 14.7% (<0. 001 mm); slit, 51.1% [9].
Different residues of plant species (trees, shrubs, and grasses)
were also sampled around the station, three tree species included
locust (Robinia pseudoacacia), simon poplar (Populus.simonii Carr)
and elm (Ulmus pumila L.); three shrubs were peashrub (Caragana
korshinskii Kom.), Sea buckthorn (Hippophae rhammoides L.), wild
peach (Prunus davidiana (Carr.) Franch); four grasses were Stipa
bungeana (Stipa bungeana), old world bluestem (Bothriochloa
ischaemum (L.) Keng), Milkvetch (Astragalus adsurgens Pall), alfalfa
(Medicago sativa L.).
These plants are the native species in the region. When sampling, the foliar were taken from trees and shrubs, and; the aboveground were taken for grasses. And the samples were washed, and
air-dried, and then dried in 60 C oven, and sieved into 0.51.0 mm
for use. And the contents of total C, N and lignin in the residues are
presented in Table 1.
2.2. Incubation experiment
Before the incubation experiment, air-dried soil was pre-incubated at 70% of eld capacity for a week to activate the soil microbial activity. Moisture soil equal to 600 g of air-dried soil was
mixed with 1% of different residues of plant, and added into an
incubation box, and incubated in 28 C incubator. Soil without
addition of plant residues was as control. Three replicates for each
treatment. Sub-sample was taken from each incubation box at 0, 1,
3, 6, 10, 16, 24, 34, 46, and 60 days of incubation to determine contents of soil microbial biomass carbon and nitrogen. Water was
added by weight method every 3 days to keep the soil moisture
stable during the incubation.
2.3. Determination methods
Soil moisture was determined with oven drying method, and
nutrients in soil and plant were determined by conventional method [10]. The lignin was determined with iodometry method [11].
Soil microbial biomass carbon and nitrogen were determined with
fumigation and extraction method [12].
Briey, soil sample was fumigated with CH3OH-free CHCl3 for
24 h at 25 C (fumigated samples) to destroy cell membranes of
soil microorganisms (unfumigated soil as the control). After removal of the CHCl3 vapor, the fumigated and unfumigated samples
were extracted with 0.5 M K2SO4 solution. Total C in the ltrate

was determined with TOC analyzer (Phoenix 8000); and total

soluble N in the ltrate was oxidized with alkaline persulfate oxidation, then measured by dual-wavelength ultraviolet spectrophotometer [13]. Both SMBC and SMBN were calculated as the
differences in ltrates between fumigated and unfumigated soil.
A kC and kN factor for SMBC and SMBN were both 0.45 [12].
2.4. Statistics
Differences between different treatments were evaluated with
SAS (8.0) for each examined parameter using the Duncan honestly
signicant difference test at 95% signicance level.
3. Results
3.1. Chemical compositions of residues of different plant species
The coefcients of variation (CV) of N contents in the plant residues was as high as 47.4%, and alfalfa with the highest N content
32.78 g kg 1, old world bluestem with the lowest N 6.61 g kg 1
(Table 1). Compared to the N contents in plant residues, the variation of C contents in plant residues was relatively low (CV = 5.85%);
C content in S. bungeana was the highest (499.46 g kg 1), and simon poplar was the lowest (414.44 g kg 1). Therefore, it resulted
in high variation of C/N ratio of plant residues (CV = 65.8%); the
highest was old world bluestem (C/N = 65.49), the lowest was alfalfa C/N 14.19. The C/N ratio of simon poplar was the highest
among the three trees species, next were locust and elm. The variation of C/N ratio of three shrubs was small, with average of 15.64.
For grasses, the C/N ratios of S. bungeana and old world bluestem
were higher than alfalfa and Milkvetch.
The average lignin contents in the residues was 27.11%, and the
highest was simon poplar (30.89%), the lowest was milkvetch
(24.10%), with CV 8.2%. The lignin contents of simon poplar in three
tree species was the highest, and next were locust and elm; the
lignin contents in three shrub species followed the order: peashrub > Seabuckthorn > wild peach. For grasses the lignin contents
of S. bungeana and old world bluestem were higher than alfalfa and
Milkvetch.
3.2. Mineral N in different treatments
The differences of mineral N in soil with and without addition of
plant residues during the incubation indicate the net mineralization of the residues. Positive value indicated the release of N from
the residues, and negative value indicated the immobilization of N.
Additions of simon poplar and locust resulted in net immobilization of N during the incubation, and N immobilization by simon

Table 1
Chemical component properties of the plant residues.
Plant residues

TOC (g kg

Total N (g kg

C/N ratios

Lignin (%)

Trees

A
B
C

Ulmus pumila L.
Populus.simonii Carr
Robinia pseudoacacia

422.58
414.44
437.73

25.36
9.02
14.92

16.66
45.97
29.33

25.20
30.89
27.78

Shrubs

D
E
F

Caragana korshinskii Kom

Hippophae rhammoides L.
Prunus davidiana (Carr.) Franch

466.85
464.70
458.53

30.95
29.71
28.29

15.08
15.64
16.21

29.59
27.14
25.76

Grasses

G
H
I
J

Stipa bungeana
Bothriochloa ischaemum (L.) Keng
Astragalus adsurgens Pall
Medicago sativa L.

499.46
432.69
427.93
464.98

9.86
6.61
27.01
32.78

50.67
65.49
15.84
14.19

27.54
28.61
24.10
24.47

Average

448.99

21.45

28.51

27.11

Coefcient of variation (%)

5.84

47.43

65.81

8.23

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J.-B. Zhou et al. / Acta Ecologica Sinica 30 (2010) 123128

140

140

120

120

100

100

Mineralized N (mgkg -1)

Mineralized N (mgkg -1)

80
60
40
20
0
-20

10

20

30

40

50

60

-40
-60

80
60
40
20
0

10

20

30

40

50

60

-20
-40
-60

Incubation time (d)

140

Incubation time (d)

I

Mineralized N (mgkg -1)

120
100
80
60
40
20
0
-20

10

20

30

40

50

60

-40
-60

Incubation time (d)

Fig. 1. Nitrogen mineralization from the different plant residues during the incubation. (A) Ulmus pumila L., (B) Robinia pseudoacacia, (C) Populus.simonii Carr, (D) Caragana
korshinskii Kom, (E) Hippophae rhammoides L., (F) Prunus davidiana (Carr.) Franch, (G) Stipa bungeana, (H) Bothriochloa ischaemum (L.) Keng, (I) Astragalus adsurgens Pall, (J)
Medicago sativa L. (the same below).

poplar was higher than locust. For elm, net N immobilization occurred during the early 20 days of the incubation, then N release
occurred (Fig. 1a).
There were signicant differences in N release from residues of
three shrub species during the incubation (Fig. 1b). For peashrub,
net N mineralization was positive, indicating the release of N from
the residues during the entire incubation. For Seabuckthorn and
wild peach, N immobilization occurred before 15 days and 34 days
of incubation, respectively. After that, the immobilized N was
released.
During the incubation, addition of S. bungeana and old world
bluestem resulted in the net immobilization of N (Fig. 1c), and N
immobilized by S. bungeana was higher than old world bluestem.
Addition of alfalfa and milkvetch resulted in net releases of N during incubation, and N released from alfalfa was signicantly higher
than milkvetch (P < 0.01).

3.3. Soil microbial biomass C (SMBC) in different treatments

Compared to the treatment without addition of residues, addition of different plant residues increased the contents of SMBC during the incubation (Fig. 2). During the initial stage of incubation,
SMBC increased sharply, and peaked at 6 days of incubation, and
then declined gradually.
The increases of SMBC in elm treatment during the initial stage
was higher than simon poplar and locust (Fig. 2a); and as the incubation continuing, the SMBC in elm treatment decreased; and it
was signicantly higher than simon poplar and locust treatment
during the rst 30 days of the incubation. The SMBC contents in simon poplar and locust followed a same trend during the incuba-

tion; at the end of incubation, the average of SMBC of locust

treatment was higher than simon poplar treatment (P < 0.05).
For three shrub species (Fig. 2b), SMBC contents of peashrub
and wild peach treatments were signicantly higher than Seabuckthorn (P < 0.05) during the incubation. For three grass species
(Fig. 2c), SMBC contents of milkvetch and alfalfa followed a same
trend, in the early 30 days of the incubation, SMBC in these treatments were signicantly higher than S. bungeana and old world
bluestem (P < 0.05).
At the end of incubation, SMBN contents in soils added with different plant species was shrubs (average, 269.68 mg kg 1) > grass
(244.67 mg kg 1) > trees (231.26 mg kg 1), and the difference was
statistically signicant.

3.4. Soil microbial biomass N (SMBN) in different treatments

Compared to the treatment without addition of the residues,
addition of different plant residues increased the contents of SMBN
during the incubation (Fig. 3). As the continuing of the incubation,
SMBN declined gradually, but was still higher than treatment without addition of residues.
SMBN content in elm treatment was higher than simon poplar
and locust treatments (Fig. 3a). During the rst 35 days of the incubation, SMBN in locust treatment was higher than simon poplar
treatment (P < 0.05). During the late stage of the incubation, the
differences between different residues of three tree species became
smaller. The SMBN contents in peashrub and wild peach treatments were signicantly higher than Seabuckthorn treatment
(P < 0.05). At the end of the incubation, the SMBN contents in peashrub and wild peach treatments were 68.8% and 49.2% higher

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J.-B. Zhou et al. / Acta Ecologica Sinica 30 (2010) 123128

b
700
CK

700

Soil microbial biomass C

(mgkg-1)

600
500
400
300
200
100
0

600

CK

500
400
300
200
100
0

10

20
30
40
Incubation time (d)

Soil microbial biomass C (mgkg -1)

Soil microbial biomass C (mgkg-1)

50

60

10

20
30
40
50
Incubation time (d)

60

700
600

CK

500
400
300
200
100
0
0

10

20

30

40

50

60

Incubation time (d)

Fig. 2. Changes of soil microbial biomass C under different treatments during the incubation. CK = soil (the same below).

Soil microbial biomass N (mgkg -1)

b
140
120
100
80

CK

60
40
20
0

10

20
30
40
Incubatin time (d)

Soil microbial biomass N (mgkg -1)

Soil microbial biomass N (mgkg -1)

50

60

140
120
100

CK

80
60
40
20
0
0

10

20
30
40
Incubatin time (d)

140
CK
H
J

120
100

G
I

80
60
40
20
0

10

20
30
40
Incubatin time (d)

50

60

Fig. 3. Changes of soil microbial biomass N under different treatments during the incubation.

50

60

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J.-B. Zhou et al. / Acta Ecologica Sinica 30 (2010) 123128

Table 2
Correlation analysis among soil microbial biomass carbon and nitrogen after 60-day incubation, mineralized N from plant residues and its chemical components.
SMBC
SMBC

SMBN

0.773** (n = 100)

**

N-mineralized
from residues

C/N ratio

0.065 (n = 100)

0.325** (n = 100)

C/N ratio

0.687* (n = 10)

0.683* (n = 10)

0.712* (n = 10)

0.583 (n = 10)

0.488 (n = 10)

0.565 (n = 10)

0.712 (n = 10)

Lignin of
residues

Organic-C of
residues

Total-N of
residues

N-mineralized

Lignin of residues

SMBN

Organic-C of residues

0.310 (n = 10)

0.404 (n = 10)

0.254 (n = 10)

0.069 (n = 10)

0.071 (n = 10)

Total-N of residues

0.807** (n = 10)

0.800** (n = 10)

0.841** (n = 10)

0.946** (n = 10)

0.578 (n = 10)

0.202 (n = 10)

Indicate the signicant difference at P < 0.05.

Indicate the signicant difference at P < 0.01.

than Seabuckthorn treatment, respectively. During most of incubation, the SMBN contents in alfalfa and milkvetch treatments were
higher than S. bungeana and old world bluestem treatments
(Fig. 3c); and SMBN of alfalfa was higher than other grass species.
At the end of 60 days incubation, SMBN in soil added with the
different residues of plant species followed the order of shrubs (average, 53.44 mg kg 1) > grass(43.28 mg kg 1) > tree(30.37 mg kg 1),
and the differences between each species was statistically
signicant.
3.5. Correlationships between SMBC, SMBN and mineralized N and
chemical composition of residues
There were signicantly positive correlationships between total
N in the residues and SMBC, SMBN and mineralized N and chemical
composition of residues (Table 2); and negative correlationships
was found between total N in the residues and C/N ratio (P <
0.05), and the correlationship between total N and lignin was not
statistically signicant. SMBN also had a positive correlationships
with mineralized N from the residues and SMBC (P < 0.01), and
SMBCs correlationships with mineralized N was not statistically
signicant.
4. Discussion
There are many researches to study the N release from different
plant residues [9,14,15]. Addition of residues with higher C/N ratio
resulted in N immobilization due to high energy for microbes; on
the contrast, addition of residues with low C/N ratio (e.g., legume)
increased N released from the residues [16]. However, the study on
the N release from different plant residues of Loess Plateau is limited. This study showed that after addition of residues of alfalfa,
milkvetch, and peashrub, N released from these residues since
the initial of the incubation. For elm, Sea buckthorn and wild peach
treatments, N immobilization occurred during the rst 14 to
34 days of incubation, then released. And for simon poplar, locust,
S. bungeana and old world bluestem treatments, N from these residues did not release during the entire incubation, mineral N in soil
was immobilized. There was positive correlationship between the
N released from residues and their total N content; and negatively
correlationship with the C/N ratio (Table 2). It indicates that the N
released from the residues is closely related to total N and C/N ratio
of the residues.
Addition of the plant residues signicantly increased the contents of SMBC, SMBN in soil. It is consistent with the other studies
[1,1719]. This is related to the increasing activity of microbes by
addition of organic matter. However, different residues of plant
species had various effects on the contents of SMBC, SMBN in soil.
After the incubation, generally, addition of the residues with higher

N content and low C/N ratio resulted in higher contents of SMBC,

SMBN in soil than the residues with lower N content and higher
C/N ratio. For example, at the end of incubation, the SMBC, SMBN
in soil added the alfalfa (with the lowest C/N ratio, 14.19) were
higher than the old world bluestem residues (with the highest C/
N ratio, 65.47). There was negatively correlationship between the
SMBC, SMBN in soil and total N and C/N ratio of plant residues (Table 2). It indicates that the C/N ratio of plant residues was the main
reason for the differences in SMBC, SMBN in soil [20]. This is related to the higher decomposition rate of low C/N ratio residue
after addition into soil [21].
The C/N ratios of three shrubs (Sea buckthorn, peashrub and
wild peach) was similar (C/N, 1516). However, after addition of
these residues to soil, there were signicant differences in SMBC,
SMBN; and N released from these residues were also signicant
differences (Fig. 3b). It indicates that the C/N ratio of residues is
not only one factor affecting their decomposition; and other chemical compositions of residues, such as cellulose, hemicelluloses, and
lignin, etc., also affect their decomposition [2225]. Therefore, it is
needed to study the relationship between the different chemical
compositions of residues from the Loess Plateau and their decomposition rates.
Water stress is known as a limited factor for reestablishing the
vegetation on the Loess Plateau. Therefore, different types of plants
including trees, shrubs, or grasses are chosen at the basis of water
resource in different regions [2628]. This study indicated that different residues of plant species had different chemical properties
(Table 1), and N releasing characteristic, and their addition on
SMBC, SMBN were also different. Undoubtedly, these differences
(especially in N) have great effects on transformations of organic
matter and energy ow in soilplant ecosystem. Therefore, it is
suggested that during reestablishing the vegetation on the Loess
Plateau, the characteristics of C and N decomposition from different residues of plant is needed to consider to adjusting C and N cycles in soil ecosystem.
In this study, we used the incubation method to evaluate the effects of addition of plant residues from the Loess Plateau on mineral N, SMBC, SMBN in soil. It is clear that the transformation of
plant residues in soil in the eld is affected by environmental factor, such as soil moisture, temperature, etc. Therefore, long term
eld study is needed to study the decomposition of plant residues
in soil and their effects on soil fertility.
Acknowledgements
This research was supported by the National Natural Science
Foundation of China (No. 40571087) and National Technology
R&D Pillar Program in the 11th Five Year Plan of China
(2007BAD89B02).

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J.-B. Zhou et al. / Acta Ecologica Sinica 30 (2010) 123128

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