Journal of Integrative Agriculture 2022, 21(5): 1488–1500

Available online at www.sciencedirect.com

ScienceDirect

RESEARCH ARTICLE

Increasing soil microbial biomass nitrogen in crop rotation systems

by improving nitrogen resources under nitrogen application

XING Ting-ting1, 2, CAI An-dong3, LU Chang-ai1, YE Hong-ling1, 2, WU Hong-liang1, HUAI Sheng-chang1,

WANG Jin-yu1, XU Ming-gang1, LIN Qi-mei3

1
Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences/National Engineering
Laboratory for Improving Quality of Arable Land, Beijing 100081, P.R.China
2
College of Land Science and Technology, China Agricultural University, Beijing 100094, P.R.China 
3
Institute of Environment and Sustainable Development in Agriculture, Chinese Academy of Agricultural Sciences, Beijing 100081,
P.R.China

Abstract
Soil microbial biomass nitrogen (MBN) contains the largest proportion of biologically active nitrogen (N) in soil, and is
considered as a crucial participant in soil N cycling. Agronomic management practices such as crop rotation and mono-
cropping systems, dramatically affect MBN in agroecosystems. However, the influence of crop rotation and mono-
cropping in agroecosystems on MBN remains unclear. A meta-analysis based on 203 published studies was conducted
to quantify the effect of crop rotation and mono-cropping systems on MBN under synthetic N fertilizer application. The
analysis showed that crop rotation significantly stimulated the response ratio (RR) of MBN to N fertilization and this
parameter reached the highest levels in upland-fallow rotations. Upland mono-cropping did not change the RR of MBN
to N application, however, the RR of MBN to N application in paddy mono-cropping increased. The difference between
crop rotation and mono-cropping systems appeared to be due to the various cropping management scenarios, and the
pattern, rate and duration of N addition. Crop rotation systems led to a more positive effect on soil total N (TN) and a
smaller reduction in soil pH than mono-cropping systems. The RR of MBN to N application was positively correlated with
the RR of mineral N only in crop rotation systems and with the RR of soil pH only in mono-cropping systems. Combining
the results of Random Forest (RF) model and structural equation model showed that the predominant driving factors of
MBN changes in crop rotation systems were soil mineral N and TN, while in mono-cropping systems the main driving
factor was soil pH. Overall, our study indicates that crop rotation can be an effective way to enhance MBN by improving
soil N resources, which promote the resistance of MBN to low pH induced by intensive synthetic N fertilizer application.

Keywords: microbial biomass nitrogen, crop rotation systems, mono-cropping systems, synthetic nitrogen  fertilizer,
meta-analysis

Received 29 December, 2020 Accepted 2 March, 2021

XING Ting-ting, E-mail: 15947039328 @163.com; Correspondence 1. Introduction
LU Chang-ai, E-mail: luchangai@caas.cn
© 2022 CAAS. Published by Elsevier B.V. This is an open Soil microbes are a crucial component in soil organic
access article under the CC BY-NC-ND license (http://
creativecommons.org/licenses/by-nc-nd/4.0/).
matter decomposition and sequestration as well as
doi: 10.1016/S2095-3119(21)63673-0 carbon and nitrogen (N) cycling (Manzoni and Porporato
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500
2009; Gessner et al. 2010). Soil microbial biomass N
(MBN) plays a critical role in maintaining soil fertility
and is considered as a biologically active N pool in soils
(Deng et al. 2000). Despite its small size (0.5–15.3% of
total soil N), MBN is important in the conversion between
soil organic and inorganic N pools and hence critical to
regulating plant nutrient uptake (Anderson and Domsch
2006). The size of the MBN pool is affected by changes
in agricultural management practices and could indicate
early changes in soil N stability due to its high sensitivity
to changes in the soil environment (Li et al. 2018). A large
number of studies have found an increase in MBN under
synthetic N fertilizer application (Liu et al. 2010; Gong
et al. 2011; Liang et al. 2011; Qiu et al. 2016; Shahid
et al. 2017), however, it is still unknown whether MBN is
influenced by crop rotation management.
Crop rotation and mono-cropping are major cropping
management systems used in agroecosystems. Crop
rotation is effective in mitigating weed, insect and pathogen
pressure due to the improved richness of residual roots
and litter in cropping soils which may improve crop yields
(Smith et al. 2008). Moreover, enhanced crop diversity
in crop rotations may increase MBN (Moore et al. 2000;
Mcdaniel et al. 2014; Peralta et al. 2018) by improving
soil microbial community diversity, growth efficiency,
and enzyme activity, which leads to stimulated microbial
resistance to environmental changes (Li et al. 2009;
Mcdaniel et al. 2014; Zhang et al. 2015). In contrast,
mono-cropping practices deplete soil nutrients, reduce
bacterial abundance and community diversity due to the
autotoxicity of root exudates, reduce nutrient utilization
and microbial resistance to environmental changes
(Lithourgidis et al. 2006; Li et al. 2019). Application of
synthetic N fertilizers indirectly influences soil microbes
through affecting soil fertility in agroecosystems.
Consequently, how MBN changes in crop rotation and
mono-cropping systems following N fertilizer application
and whether MBN is influenced by N fertilizer application
pattern, duration and rate need to be clarified.
Soil properties are influenced by cropping management
system and altered soil properties may drive changes
in MBN (Marais et al. 2012). Crop rotation practices
enhance soil organic matter input derived from various
crop residues and thus improve soil quality and structural
stability (Mcdaniel et al. 2014; Peralta et al. 2018).
For instance, soil organic carbon (SOC), total N (TN)
content and pH are higher in crop rotation than in mono-
cropping systems (Lithourgidis et al. 2006; Li et al. 2019).
Furthermore, application of synthetic N fertilizers in both
crop rotation and mono-cropping systems enhance crop
yield and plant C inputs into the soil, which leads to the
increase in microbial biomass due to alleviation of C
1489
and N limitation (Zhou et al. 2002; Fierer et al. 2009;
Chen et al. 2018). On the other hand, application of N
fertilizers may reduce soil pH (Zhang et al. 2009; Guo
et al. 2010; Tian and Niu 2015), resulting in stimulation
of leaching of Ca 2+, Mg 2+ and Na + which are required
for microbial growth, and also mobilization of Al3+ which
is harmful to microorganisms (Treseder 2008; Lu et al.
2011; Chen et al. 2015; Deng et al. 2017). Crop rotation
practices have positive effects on microbial diversity and
soil fauna and microbial abundance, which may improve
the resistance of microbes to low soil pH compared with
mono-cropping systems (Zhang et al. 2015). On the
other hand, increasing crop diversity through crop rotation
improves SOC and TN, which causes only minor changes
in soil pH due to enhanced soil acid-buffering capacity
(Mcdaniel et al. 2014). MBN is affected by soil properties,
but it is unclear whether different soil factors control the
response of MBN to N fertilizer application in crop rotation
and mono-cropping systems.
To date, two meta-analyses have compared the
response of soil MBN to synthetic N fertilizer application
in agroecosystems (Lu et al. 2011; Zhang Q et al. 2017).
The study of Zhang Q et al. (2017) focused on Chinese
farmland while that of Lu et al. (2011) compiled data from
only 18 studies including MBN (from 1985 to 2007). Both
analyses lack data on the MBN response to N fertilizer
application in crop rotation and mono-cropping systems
and information on the mechanisms involved. In the last
10 years, a number of field studies have focused on the
effect of synthetic N fertilizer application on MBN under
various cropping systems. We performed a comprehensive
meta-analysis to integrate 203 relevant published papers
to: 1) characterize how soil MBN differs in crop rotation
and mono-cropping systems and 2) clarify whether there
are different driving factors influencing MBN in the two
management systems. Our hypotheses included: (1) crop
rotation practices lead to higher soil MBN levels than
mono-cropping practices, and (2) improvements in soil N
resources due to synthetic N fertilizer application would
drive changes in MBN in crop rotation systems.
2. Materials and methods
2.1. Data collection
For the current study, “soil microbial N”, “cropland”
or “farmland”, “crop rotation”, “mono-cropping” or
“monoculture” and either “N fertilizer”, “N fertilization”,
“N input”, “N addition”, “N application”, or “N enrichment”
were used as search items for the China National
Knowledge Infrastructure (CNKI) (http://www.cnki.net/)
and Web of Science (http://apps.webofnowledge.com/)
1490 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500
in January 2018. Overall, 203 papers including 144 sites
were selected (Appendix A). Criteria for including studies
in the analysis were as follows: (1) studies must have
been farm-based or controlled laboratory experiments;
(2) the study must have included a paired synthetic N
fertilizer group and non-fertilized control group; and (3)
all treatments must have been replicated and results
must have been presented with measures of variability
including standard deviations (SDs) or standard error
(SEs). If the SD was not provided, an estimate was
derived as SD=mean/10 (Luo et al. 2006). (4) If a study
was conducted over multiple years, the sampling interval
must have been less than five years. Data protocols
used for the selected studies were as follows. If the study
contained results from different experimental sites, results
of different sites were included as independent studies.
Our data only included one growing season; if a paper
included different plant growth stages, the arithmetic
mean of each growth period was included in the database.
If treatments in one paper included experimental results
of conventional tillage and no-tillage, only experimental
results of conventional tillage were included. If data in
the paper were represented as graphs or other figures,
means and standard deviations were extracted using
GetData Graph Digitizer 2.26 Software (http://www.
getdata-graph-digitizer.com/index.php).
In the selected studies, MBN was determined using a
chloroform fumigation extraction or chloroform fumigation
culture technique. The methods used for determining
other soil physical and chemical properties given in
the studies (soil pH, TN, the ratio of SOC to TN (C/N),
dissoluble organic N (DON), ammonium N, nitrate N,
mineral N ((ammonium N + nitrate N)) are presented in
the original studies but were not included here.
The assembled database contained latitudinal and
longitudinal information, soil properties (soil type, soil pH),
climate information (mean annual temperature (MAT) and
mean annual precipitation (MAP), cropping system, crop
type, type of synthetic N fertilizer application (application
alone or as a mixture with other fertilizers), application
rate, fertilization duration, and form of N fertilizer. Soil
type and texture were classified according to the US Soil
Classification System (Soil Survey Staff 2006). Mean
annual temperature (MAT) was divided into groups
consisting of <5, 5–10, 10–20 or >20°C, mean annual
precipitation (MAP) was divided into groups of <500,
500–1 000, 1 000–1 500, or >1 500 mm. Crop types and
crop rotation systems were identified from each original
study and compiled into the database (Table  1). The
type of synthetic N fertilizer application was grouped into
N application alone (N), N combined with phosphorus
(NP), and N combined with phosphorus and potassium
(NPK). Mean annual mineral N fertilizer application rate
was grouped into <100, 100–300, 300–500, or >500 kg
ha–1 yr–1. Synthetic N fertilizer application duration was
divided into <1, 1–10, 11–20, 21–30, or >30 years. Form
of N fertilizer was categorized as urea, NH4+, NO3–, or
NH4++NO3– (Appendix B).
2.2. Meta-data analysis
MetaWin 2.1 Software (Rosenberg et al. 2000) was used
to assess the effect of N fertilizer application on MBN.
The response ratio (RR) was calculated as the ratio of
the natural logarithm of a given variable in the treatment
group to that in the control group (eq. (1)) (Hedges et al.
1999):
RR=ln(X /Xt c)=ln(Xt )–ln(Xc)  (1)
where X t and X c are the mean value of the variable X of
the treatment group and the control group, respectively.
The variance of effect size was calculated as:
St2 Sc2
V= + (2)
ntXt2 ncXc2 
where n t and n c are the number of samples in the
treatment and control group, respectively, and St and Sc
represent the standard deviations of the treatment and
control group, respectively. Weight was calculated as:
Wij =1/V  (3)
where Wij is the weighting factor for the ith level and the
jth pair and V is a variance.
The weighted response ratio (RR++) was used to represent
overall response of the treatment group compared to the
control group and calculated as:
m k
∑ ∑ WijRR
i=1 j=1
RR++= m k  (4)
∑ ∑ Wij
i=1 j=1
where m is the number of compared groups, and k is the
number of comparisons in the corresponding groups. The
standard error of RR++ (S(RR++)) was calculated as:
1
S(RR++)= m k
 (5)
∑ ∑ Wij
i=1 j=1
If data were not normally distributed or had less
than 20 data points, the bootstrap method was used to
calculate the 97.5 percentile of RR++ over 5 000 iterations
(Zhou et al. 2014). If data were normally distributed or
had greater than 20 data points, the 95% confidence
interval (95% CI) of RR++ was used and calculated as:
95% CI= RR++ ±1.96×S(RR++) (6)
If the RR ++ confidence interval did not overlap the
abscissa zero point, means were considered significantly
different for the control and treatment groups (Gurevitch
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500 1491

Table 1 A list of crop type and cropping system and each group tested for the significance of MBN in the meta-analysis
Variable Group Explanation
Crop type Cereal Maize, wheat, sorghum, oats, barley, rice
Legume Soybeans, peanuts, cowpeas
Tuber Potato, sweet potato
Vegetable Tomato, celery, eggplant, cucumber, spinach
Other crops Tobacco, sugar cane, rape, cotton, alfalfa, strawberry
Cropping system
Rotation system Upland crop rotation system Wheat–maize
Maize–maize–legume
Maize–legume–wheat
Maize–legume
Maize–wheat-legume
Legume–wheat–maize
Legume–legume–legume–legume–wheat
Maize–legume–maize
Wheat–legume–maize
Tomato–celery
Wheat–potato
Potato–wheat–green manure
Tomato–tomato–tomato–eggplant
Sweet potato–rape
Wheat–rape
Upland crop-fallow Wheat–fallow
rotation Maize–maize–fallow
Tomato–fallow
Paddy crop-fallow rotation Rice–rice–fallow
Upland-paddy crop Rice–rape
rotation Rice–rice–rape
Rice–wheat–jute
Rice–wheat–legume
Hordeum–rice–rice
Rice–maize
Wheat–rice
Rice–rice–wheat
Mono-cropping system Upland mono-cropping Upland crop
Paddy mono-cropping Paddy crop

and Hedges 1999). RR++ was converted to a percentage
(%) using the following equation to assess percent
change (%):
Percentage (%)=(eRR++–1)×100
2.3. Data analysis
It was necessary to conduct a bias test for collected
documents to remove articles which exhibited
characteristics of publication bias before the analysis
(Deng et al. 2015). Results of the publication bias
assessment demonstrated that the data were independent
and there was no publication bias in regards to MBN
(Appendix C). Because a large proportion of the data was
not normally distributed (Appendix D), a boot-strapping
method was selected to meet requirements of normally
distributed data (Jian et al. 2016). The Chi-square test
was used to assess differences in heterogeneity among
groups at P<0.05. The Chi-square test was used to
perform between-group (QB) and within-group (QW)
heterogeneity tests (Treseder 2008).
Random Forest (RF) is a regression model that
combines multiple decision trees to improve prediction
accuracy (Shahbazi et al. 2017; Zhang H et al. 2017) and
was used to identify factors impacting MBN response
following application of synthetic N fertilizers. The RF
method independently considers the importance of each
influencing factor, interdependence of influencing factors,
and ranks influencing factors according to their importance
(Strobl et al. 2007; Shahbazi et al. 2017). In the RF
model, the importance score is calculated by out-of-bag
(OBB) data to calculate the mean square error (MSE),
and the OBB data are not involved in the tree construction
process of random data subsets (Zhang H et al. 2017).
The final value of the importance score is determined by
the sum of MSE calculated across all trees. The larger
the value is, the more important the factor is (Shahbazi
et al. 2017). The RF model was implemented using the
1492 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500

package “Random forest” in R (R Core Team 2018). The
“Miss Forest” function was used to impute missing values,
and then to determine the importance of N addition rate
(kg N ha–1 yr–1), N duration (yr), climatic factors and soil
physical and chemical properties on the response ratio of
MBN to N fertilization.
A piecewise structural equation model (SEM) (Lefcheck
and Duffy 2015) was used to quantify the relationships
among MBN, soil properties, and fertilizer managements
(N fertilizer addition rate and duration). The piecewise
SEM is translated to multiple linear mixed relationships
and evaluated individually to test the direct and indirect
effects (Lefcheck 2016). Shipley’s d-separation test of
generalized causal path analysis depended on a set of
regressions to find whether any paths are missing from the
model (Shipley 2009). It used akaike information criterion
(AIC) to compare nested models with small sample size
(Shipley 2013), with the traditional variance covariance-
based SEM (http://jonlefcheck.net/2014/07/06/piecewise-
structural-equation-modeling-in-ecological-research/). We
constructed the piecewise SEM based on the schematic
diagram in the Appendix E, which was implemented in the
R package ‘piecewiseSEM’ (https://github.com/jslefche/
piecewiseSEM) and reported the standardized coefficient
for each path from each component model.
3. Results
3.1. Variation in MBN between crop rotation and
mono-cropping systems
Synthetic N fertilizer application increased soil MBN by
38.72% in crop rotation systems compared to 12.5% in
mono-cropping. In addition, the upland, upland-fallow,
and upland-paddy rotations had a positive effect on the
response of MBN to N fertilizer application with the effect
being greatest for the upland-fallow rotation (124.16%,
P<0.01; Fig. 1). Upland mono-cropping had no effect on
MBN but paddy mono-cropping increased the response
ratios (RRs) of MBN to N fertilizer application by 28.16%
among all kinds of mono-cropping. The MBN/TN ratio
was significantly enhanced by application of N fertilizer
in crop rotation systems with the positive effect being
greatest in the upland crop-fallow rotation (189.85%)
(Appendix F).
Differences in MBN between crop rotation and mono-
cropping systems varied with different synthetic N fertilizer
management. MBN was greater in crop rotation systems
than in mono-cropping, when N fertilizers were applied
alone, synthetic N fertilizer addition rate was less than
300 kg ha–1 yr–1, or when the duration of application was
less than one year (Fig. 2).
3.2. Effects of crop rotation and mono-cropping
systems on soil chemical properties
Soil mineral N increased by 61.67 and 63.17% in
response to N fertilizer application in crop rotation and
mono-cropping systems, respectively (Fig.  3). The
positive effect on the response of TN to N fertilizer
application in crop rotation systems (21.74%) was higher
than that in mono-cropping systems (7.56%). Soil C/N
was found to be significantly enhanced (14.16%) in
response to N fertilization in crop rotation systems, but not

Crop rotation Mono-cropping

QB=29, QW=778, P<0.01

Crop rotation a 339
Mono-cropping b 91

QB=128, QW=804, P<0.01

Upland rotation b 201

Upland mono-cropping c 54

Upland-paddy rotation b 104

Paddy mono-cropping b 41
a
Upland-fallow rotation 30

Paddy-fallow rotation bc 14

–40 0 40 80 120 160 200

Percentage change (%)

Fig. 1 Effects of crop rotation and mono-cropping on soil MBN under synthetic N fertilizer application. Error bars show SD. The
sample size (n) is displayed beside each bar. Cropping systems with different lowercase letters are significantly different at P<0.05.
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500 1493

Crop rotation Mono-cropping

N N Duration a 28
addition a 10 addition a 122 >30
a 14
NPK type 300–500 rate
a 39 a 9 a 80
21–30
b 21
a 45 a 168 a 86
NP 100–300 11–20
a 42
a 13 b 76
a 121
1–10
a 142 a 19 a 8
N <100 a 29
b 41 b 9 <1
b 10

-40 0 40 80 120 -40 0 40 80 120 -40 0 40 80 120

Percentage change (%)

Fig. 2 Crop rotation and mono-cropping effects on soil microbial biomass nitrogen under different synthetic N fertilizer type (NPK, NP
or N alone), rate (kg ha–1 yr–1) and duration (number of years of application). Error bars show SD. The sample size (n) is displayed
beside each bar. Different lowercase letters indicate significant differences between different N fertilizer managements at P<0.05.

changed in mono-cropping systems. N fertilizer increased
soil DON, nitrate N and ammonium N by 27.05, 92.13
and 79.36%, respectively, in crop rotation systems and by
33.16, 50.43 and 45.50%, respectively, in mono-cropping
systems. In addition, crop rotation systems (–2.87%)
had a smaller reduction in N-induced changes in soil pH
compared to mono-cropping systems (–3.58%; Fig. 3).
3.3. Relationships between the response ratios
of MBN and soil factors in crop rotation and mo-
no-cropping systems
The RR of MBN had a significant positive correlation with
the RRs of TN and mineral N but had no relationship with
soil pH in crop rotation systems (Fig.  4-A–C). However,
there was a significant positive correlation between the
RRs of MBN and soil pH in mono-cropping systems
(Fig.  4-C). To clarify which factors drove the changes
in MBN in crop rotation and mono-cropping systems,
RF  model analysis was  performed. The independent
variables used in our RF model explained 58 and 57% of
the total variances in the responses of MBN to synthetic
N fertilizer application across crop rotation and mono-
cropping systems, respectively (Fig.  5-B and D). For
crop rotation systems, RRs of soil mineral N, TN and
nitrate N were the most important factors affecting the
variation in MBN response to N fertilizer application
(Fig. 5-A). RRs of DON, ammonium N, pH and duration
of N application had a small influence on MBN response
to N fertilization application. Furthermore, the RR of C/N
and rates of synthetic N fertilizer addition had a minimal
impact on MBN response (Fig. 5-A). However, for mono-
cropping systems, the most important controlling factors
for changes in MBN response to N fertilizer application
were RRs of soil pH and TN (Fig.  5-C). Consequently,
the results of RF models were accordant with the results
of regression analyses in both crop rotation and mono-
cropping systems (Fig. 4).
We also fitted a piecewise SEM in crop rotation and
mono-cropping systems, respectively, to infer the direct
and indirect effects of application of synthetic N fertilizer
(addition rate and duration) on MBN response via soil
properties. The models had Fisher’s C statistic P>0.05
and AIC=83.06 in crop rotation systems (Fig.  6-A) and
Fisher’s C statistic P>0.05 and AIC=78.92 in mono-
cropping systems (Fig. 6-B). The SEM demonstrated that
duration of synthetic N fertilizer application directly (–0.15,
standardized coefficient) and indirectly through TN (0.46,
standardized coefficient), mineral N (0.44, standardized
coefficient) and nitrate N (–0.23, standardized coefficient)
affected MBN in crop rotation systems, while the influence
of addition rate of synthetic N fertilizer on MBN was
mediated through ammonium N (–0.36, standardized
coefficient) in mono-cropping systems. Soil TN, mineral
N, nitrate N and DON directly influenced MBN response
to N fertilizer application in crop rotation systems, and
the response of soil TN and mineral N to synthetic N
fertilizer addition played a more important role (0.46 and
0.44, standardized coefficient) in affecting MBN than
did nitrate N and DON (Fig. 6-A). However, soil pH, TN
and ammonium N directly affected MBN response in
mono-cropping systems, with soil pH (0.59, standardized
1494 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500

Crop rotation Mono-cropping lead to a higher MBN than mono-cropping (Moore et al.
2000; Mcdaniel et al. 2014), however these studies
ignored the effects of crop rotation type and N fertilizer
a 121
Mineral N application. The current meta-analysis demonstrated
a 20 that upland rotations significantly stimulated the RR of
MBN to N application, but upland mono-cropping did
not change the RR of MBN (Fig. 1). This phenomenon
a 184
may be due to increased residual root and litter diversity
TN
b 55 in upland crop rotations, which stimulates soil microbial
community diversity and growth efficiency, thus ensuring
soil stability and resistance to soil environmental changes
a 123
(Moore et al. 2000; Mcdaniel et al. 2014; Peralta et al.
C/N a 47 2018). However, upland mono-cropping systems lead to
unbalanced nutrient consumption, reduced pH and SOC,
which decreases soil microbial abundance and activities
a 38 (Lithourgidis et al. 2006; Li et al. 2019). Therefore, the
DON a 21 RR of MBN in upland mono-cropping systems may have
been unable to increase largely due to relatively poor
microbial community structure. However, MBN in paddy
a 75 mono-cropping systems was increased by N fertilizer
Nitrate N application (Fig. 1). Paddy soil has a higher SOC content
a 18
than upland soil due to different Fe oxide formations
(Liu et al. 2016). Crystalline Fe oxides in paddy mono-
a 55 cropping systems are poor, which decreases rates of soil
Ammonium N a 18 organic matter decomposition under anaerobic conditions
and increases the potential of accumulating SOC due
to the large mineral surface area (Kögel-Knabner et al.
a 88 2010). SOC drives the increasing MBN content in paddy
pH mono-cropping soils because of the relatively higher
b 39
bacterial and fungal biomass than in upland mono-
–40 0 40 80 120 160 cropping soils (Liu et al. 2016). Our results showed that
Percentage change (%) upland-fallow rotation resulted in the highest increase in
the RR of MBN (Fig. 1). Secondary succession would
Fig. 3 Mean and 95% CI of weighted response ratio (RR++) happen during the fallow period and this would improve
of soil properties in crop rotation and mono-cropping systems
the diversity of plant communities, depending on seed
under synthetic N fertilizer application. Error bars show SD.
The sample size (n) of each variable is displayed beside each availability (Castro et al. 2016). Both crop diversity and
bar. Different lower-case letters indicate significant differences community composition were found to affect soil-borne
between cropping systems at the P<0.05 level.
microbial communities and improve bacteria diversity
(Zhong et al. 2007; Ceja-Navarro et al. 2010), which is of
coefficient) being more predominant than other soil benefit to MBN. Upland-fallow rotations could improve the
properties (Fig. 6-B). soil temperature, humidity and soil organic matter content,
which increases the diversity, biomass and activity of fungi
4. Discussion and bacteria and inhibits harmful microorganisms induced
by continuous mono-cropping (Li et al. 2009; Castro et al.
4.1. Effect of crop rotation and mono-cropping 2016). Consequently, upland-fallow rotations lead to the
systems on the response of soil MBN to N fertilizer highest increase in the RR of MBN to N fertilization due to
application the improvement of diversity and biomass of microbes.
Diverse N fertilizer application practices led to different
Crop rotation and mono-cropping systems might have effects on MBN in crop rotation and mono-cropping
different effects on changes in the RR of MBN to N systems (Fig. 2). Previous meta-analysis found a small
fertilizer application due to differences in crop diversity. decrease in MBN in agricultural ecosystems (Lu et al.
Previous studies have found that crop rotation systems 2011), and N fertilizer application alone inhibited MBN,
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500 1495

Crop rotation Mono-cropping

A B
4
y=0.46x+0.38 y=0.38x+0.35
R2=0.03, P<0.05 R2=0.11, P<0.01
3 y=–0.14x–0.02
y=1.27x+0.01
R2=0.14, P<0.01 R2=0.01, P=0.62
2
RR MBN

RR MBN
1

–1
–1 –0.5 0 0.5 1 1.5 RR mineral N
RR TN
C 3 D
y=0.05x+0.48
y=–0.28x+0.20
R2=0.003, P=0.67
R2=0.0014, P=0.75
2 y=0.04x–0.18
y=3.25x+0.24
R2=0.003, P=0.83
R2=0.65, P<0.01
RR MBN

RR MBN

–1
–0.5 –0.3 –0.1 0.1 –1 0 1 2
RR pH RR nitrate N

Fig. 4 Relationships between the response ratio of soil MBN (RR MBN) and soil TN (RR TN) (A), soil Mineral N (RR mineral N)
(B), soil pH (RR pH) (C) or nitrate N (RR nitrate N) (D), in different crop rotation and mono-cropping systems.

but NPK fertilizer application increased MBN in Chinese
farmland (Zhang Q et al. 2017). Relatively limited
data were available for these studies and both studies
ignored the effects of cropping management types. In
the present study, MBN was significantly increased in
crop rotation but weakly inhibited in mono-cropping
systems when N fertilizer was applied alone (Fig. 2), while
MBN was boosted in both systems when NPK fertilizer
was applied. Microbial biomass has well-constrained
C:N:P ratios (Cleveland and Liptzin 2007) and microbial
metabolic activity is mainly limited by P (Cleveland et al.
2002; Allison et al. 2008; Deng et al. 2017). Compared
to mono-cropping, crop rotation systems have been
found to return abundant and diverse crop residues
to soil, which supplied organic P to soil microbes and
stimulated the biomass of Gram positive bacteria (GP)
and fungi that favored decomposing recalcitrant nutrient
fractions (Zechmeister-Boltenstern et al. 2015). Thus, in
the present study, when N fertilizer was applied alone, it
increased MBN in crop rotation systems. In addition, crop
rotation systems had a positive effect on the RR of MBN to
N application, but mono-cropping systems did not change
the RR of MBN when N fertilizer application rate was
less than 100 kg ha–1 yr–1 and duration was less than one
year (Fig. 2). These results imply that microorganisms in
mono-cropping systems are insensitive to low N addition
rates and short-term inorganic N applications due to the
decrease in bacterial and fungal biomass (Jia et al. 2020).
A small increase in MBN may induce a significant
change in soil N cycling and its associated soil functions.
Crop rotation practices enhanced the ratio of MBN to
TN (MBN/TN), while mono-cropping systems had no
significant effect on MBN/TN (Appendix E). The MBN/
TN ratio could represent the transformation efficiency
of exogenous N input into microbial N (Joergensen and
Emmerling 2006) and MBN has been considered as an
indicator of the soil N pool (Deng et al. 2000). Therefore,
following synthetic N fertilizer application, crop rotation
1496 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500

A B
2.0
RR mineral N
RR TN 1.5 MSE=0.09
R2=0.58
RR nitrate N

Predicted RR MBN
1.0
RR DON
RR ammonium N 0.5
RR pH 0
Duration
RR C/N –0.5

Addition rate –1.0

–1.0 –0.5 0 0.5 1.0 1.5 2.0
0 10 20 30 40 50
Measured RR MBN
MSE increase (%)
C D 2.0
RR pH
RR TN 1.5 MSE=0.05

Predicted RR MBN
Addition rate R2=0.57
1.0
RR mineral N
RR C/N 0.5
RR DON
0
RR ammonium N
Duration –0.5
RR nitrate N
–1.0
0 5 10 15 20 25 30 35 –1.0 –0.5 0 0.5 1.0 1.5 2.0
MSE increase (%) Measured RR MBN

Fig. 5 Variable importance derived from the Random Forest model for response ratio of soil MBN (RR MBN) to N fertilization in
crop rotation (A) and mono-cropping (C) systems, and the performance of Random Forest models for detecting factors controlling
the RR of MBN to N fertilization in crop rotation (B) and mono-cropping (D) systems. The dashed line shows the 1:1 line.

systems, especially the upland-fallow rotation, have a
greater potential for improving both soil bio-fertility and N
sequestration than mono-cropping systems.
4.2. Driving factors affecting the response of MBN
to N in crop rotation and mono-cropping systems
As an active biological N index, MBN is affected by soil
inorganic N content and by changing soil C availability,
C:N ratio, and soil pH (Treseder 2008; Guo et al. 2010).
We found crop rotation and mono-cropping systems could
induce different responses of soil properties to synthetic N
fertilizer application, and divergent relationships between
soil properties and MBN (Figs. 3 and 4). Results from
the RF model and SEM suggested that the crucial factors
influencing the changes in MBN in crop rotation systems
were soil TN and mineral N, while in mono-cropping
systems soil pH was important (Figs. 5 and 6). A possible
explanation for these different results is that microbes in
agroecosystems are often limited by N and application
of N fertilizers can alter microbial biomass, diversity and
activities by improving N availability, SOC accumulation
from crop residues and reducing soil pH (Zhang J et al.
2017; Zhou et al. 2017). Bacteria are the dominant
microorganisms in agroecosystems and have a higher
demand for N than fungi. Bacterial dominance may be
increased by the improvement of soil N availability and
inhibited by the decreased soil pH induced by N fertilizer
application (Guo et al. 2010; Zechmeister-Boltenstern
et al. 2015). However, Acidobacteria, Actinobacteridae,
Bacteroidetes, Planctomyces and Chloroflexus can resist
low pH environmental pressure through evolution and
diffusion mechanisms (Zhang et al. 2015; Liu et al. 2016).
Moreover, GP bacteria and fungi have adaptability to low
soil pH due to their thick and interlinked peptidoglycan
cell wall (Nielsen and Ball 2015). Crop rotation leads
to a higher bacterial and fungal community abundance
and diversity (i.e., Proteobacteria, Actinobacteria and
Firmicutes) than mono-cropping, which promotes the
utilization of C and N sources and improves resistance
to decreased pH induced by application of N fertilizers
(Liu et al. 2020). Therefore, the increase in N resources
induced by application of N fertilizers controls the MBN
changes in crop rotation rather than soil pH.
We also found the negative effect of N fertilizer
application on soil pH in crop rotation systems was
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500 1497

A Fisher′s C=13.06; P=0.53; AIC=83.06

Duration RR pH

–0.48 **
**
8
RR DON –0.2 RR ammonium N

0.
0.4

38
**
3 ** 0.32**

.27 **
0.19**

–0
**
0.20

.29
–0

**
RR mineral N RR nitrate N RR TN

0.29 **

–0
.12
*
–0.23*
0.
44
**

–0.15* RR MBN 0.46**

R2=0.34

B Fisher′s C=8.92; P=0.84; AIC=78.92

Addition rate RR pH

**
8
–0.2
0.11 **

RR DON 0.27** RR mineral N

–0
.1
7 **
5 **

**
–0.31**
0.2

23
0.22

0.
**
0.47
*

RR ammonium N RR C/N RR TN

–0.59**
3 **

–0
0.3

.3
6*

RR MBN 0.59**
R2=0.55

Fig. 6 Structural equation models of synthetic N fertilizer, climate and soil chemical properties as predictors of soil MBN in crop
rotation (A) and mono-cropping (B) systems. Solid black arrows represent positive paths (P<0.05), dotted black arrows represent
negative paths (P<0.05) and dotted grey arrows represent non-significant paths (P>0.05). Numbers adjacent to arrows are
standardized path coefficients, analogous to relative regression weights, and indicative of effect size of the relationship. Overall
fit of piecewise SEM was evaluated using Shipley’s test of d-separation: Fisher’s C statistic (if P>0.05, then no paths are missing
and the model is a good fit) and Akaike information criterion (AIC). R2 values associated with endogenous variables indicate the
proportion of explained variance (adjusted R2) by relationships with other variables. *, P<0.05; **, P<0.01; Duration, synthetic N
fertilizer application duration; Addition rate, synthetic N fertilizer addition rate; Mean response ratios (RR) are presented for mineral
N, total N (TN), nitrate N, ammonium N, soil pH, dissolved organic N (DON), soil organic carbon to N ratio (C/N) and microbial
biomass N (MBN).

less than in mono-cropping systems (Fig.  3). Together systems provided higher carbon inputs and diversity
with our regression analyses (Fig.  4-C), these results of plant residues to soils, which could contain higher
showed that, compared to mono-cropping, crop rotation levels of soil organic matter and better soil structural
1498 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500
stability to improve the “buffering” capabilities provided
by soil microbes (Marais et al. 2012). The lack of crop
diversity in mono-cropping systems leads to unbalanced
nutrient consumption and as a consequence crop roots
may release H + ions because of excess uptake of
cations over anions (Tang and Rengel 2003). Secondly,
removal of alkaline products such as grain also leads to
reduced soil pH (Rengel 2011). Lower soil pH in mono-
cropping systems was not conducive to survival of most
microorganisms when compared with crop rotation
systems (Guo et al. 2010; Li et al. 2019), but increased
abundance of specific acid-tolerant microorganisms
(Shen et al. 2018). Furthermore, autotoxicity of root
exudates caused by mono-cropping systems induces
proliferation of pathogens and also reduction of
microbial activity associated with soil nitrogen cycling
(Lithourgidis et al. 2006). Consequently, mono-
cropping systems drive changes in MBN through soil
pH due to relatively poor microbial diversity. The roles
of the key bacterial and fungal communities in mono-
cropping systems still remain unknown and further
study is needed to clarify the contributions of microbial
functional groups in the response of MBN to N fertilizer
application.
5. Conclusion
We conducted a meta-analysis based on 203 articles
published globally to investigate how MBN changes in
response to N fertilization in crop rotation and mono-
cropping systems. Results of our work demonstrated
that upland rotation systems significantly stimulated
the RR of MBN to N fertilizer which reached the
highest level in upland-fallow rotation systems, while
upland mono-cropping systems did not change the
RR of MBN. However, the RR of MBN to N fertilizer
in the paddy mono-cropping systems increased
because the large mineral surface area increased the
accumulation of SOC, which increased bacterial and
fungal biomass more than in upland mono-cropping
systems. Moreover, MBN was significantly increased
in crop rotation systems but weakly inhibited in mono-
cropping systems when N fertilizer was applied alone,
but was boosted in both crop cropping systems when
NPK fertilizer was applied. Changes in MBN in crop
rotation systems were mainly stimulated by soil mineral
N and TN, but were mainly affected by the soil pH in
mono-cropping systems. These differences between
crop rotation and mono-cropping systems suggest the
need for further analysis of the microbial functional
groups and enzyme activities under a wide range of
environmental conditions. Crop rotation systems should
be considered as a means to compensate for some of
the negative effects of intensive fertilizer application
by enhancing soil microbial biomass and associated
nutrient conversion functions.
Acknowledgements
Financial supports were received from the Agro-scientific
Research in the Public Interest of China (201503122).
We thank all the researchers whose data were used in
this meta-analysis. We appreciate Prof. Li Jianwei of
Department of Agricultural and Environmental Sciences,
Tennessee State University, USA and Prof. Li Guihua of
Agricultural Resources and Regional Planning, Chinese
Academy of Agricultural Sciences for their help in this
manuscript.
Declaration of competing interest
The authors declare that they have no conflict of interest.
Appendices associated with this paper are available on
http://www.ChinaAgriSci.com/V2/En/appendix.htm
References
Allison S D, Czimczik C I, Treseder K K. 2008. Microbial activity
and soil respiration under nitrogen addition in Alaskan boreal
forest. Global Change Biology, 14, 1156–1168.
Anderson J P E, Domsch K H. 2006. Quantities of plant nutrients
in the microbial biomass of selected soils. Soil Science,
130, 211–216.
Castro H, Barrico L, Rodríguez-Echeverría S, Freitas H. 2016.
Trends in plant and soil microbial diversity associated
with Mediterranean extensive cereal–fallow rotation agro-
ecosystems. Agriculture, Ecosystems & Environment, 217,
33–40.
Ceja-Navarro J A, Rivera-Orduna F N, Patino-Zuniga L, Vila-
Sanjurjo A, Crossa J, Govaerts B, Dendooven L. 2010.
Phylogenetic and multivariate analyses to determine the
effects of different tillage and residue management practices
on soil bacterial communities. Applied and Environmental
Microbiology, 76, 3685–3691.
Chen D, Lan Z, Hu S, Bai Y. 2015. Effects of nitrogen enrichment
on belowground communities in grassland: Relative role of
soil nitrogen availability vs. soil acidification. Soil Biology
and Biochemistry, 89, 99–108.
Chen H, Li D, Zhao J, Xiao K, Wang K. 2018. Effects of nitrogen
addition on activities of soil nitrogen acquisition enzymes:
A meta-analysis. Agriculture, Ecosystems & Environment,
252, 126–131.
Cleveland C C, Liptzin D. 2007. C:N:P stoichiometry in soil:
Is there a “Redfield ratio” for the microbial biomass?
 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500
Biogeochemistry, 85, 235–252.
Cleveland C C, Townsend A R, Schmidt S K. 2002. Phosphorus
limitation of microbial processes in moist tropical forests:
Evidence from short-term laboratory incubations and field
studies. Ecosystems, 5, 680–691.
Deng Q, Hui D, Dennis S, Reddy K C. 2017. Responses
of terrestrial ecosystem phosphorus cycling to nitrogen
addition: A meta-analysis. Global Ecology and Biogeography,
26, 713–728.
Deng Q, Hui D, Luo Y, Elser J J, Wang Y P, Loladze I, Zhang
Q, Dennis S. 2015. Down-regulation of tissue N:P ratios in
terrestrial plants by elevated CO2. Ecology, 96, 3354–3362.
Deng S P, Moore J M, Tabatabai M A. 2000. Characterization
of active nitrogen pools in soils under different cropping
systems. Biology & Fertility of Soils, 32, 302–309.
Fierer N, Strickland M S, Liptzin D, Bradford M A, Cleveland
C C. 2009. Global patterns in belowground communities.
Ecology Letters, 12, 1238–1249.
Gessner M, Swan C, Dang C, McKie B, Bardgett R, Wall D,
Hättenschwiler S. 2010. Diversity meets decomposition.
Trends in Ecology & Evolution, 25, 372–380.
Gong W, Yan X, Wang J, Hu T, Gong Y. 2011. Long-term
applications of chemical and organic fertilizers on plant-
available nitrogen pools and nitrogen management index.
Biology and Fertility of Soils, 47, 767–775.
Guo J H, Liu X J, Zhang Y, Shen J L, Han W X, Zhang W F,
Christie P, Goulding K W T, Vitousek P M, Zhang F S.
2010. Significant acidification in major Chinese croplands.
Science, 327, 1008–1010.
Gurevitch J, Hedges L V. 1999. Statistical issues in ecological
meta-analyses. Ecology, 80, 1142–1149.
Hedges L V, Gurevitch J, Curtis P S. 1999. The meta-analysis
of response ratios in experimental ecology. Ecology, 80,
1150–1156.
Jia X Y, Zhong Y Q W, Liu J, Zhu G Y, Shangguan Z P, Yan W
M. 2020. Effects of nitrogen enrichment on soil microbial
characteristics: From biomass to enzyme activities.
Geoderma, 366, 114256.
Jian S, Li J, Chen J, Wang, G, Mayes M A, Dzantor K E, Hui
D, Luo Y. 2016. Soil extracellular enzyme activities, soil
carbon and nitrogen storage under nitrogen fertilization: A
meta-analysis. Soil Biology and Biochemistry, 101, 32–43.
Joergensen R G, Emmerling C. 2006. Methods for evaluating
human impact on soil microorganisms based on their
activity, biomass, and diversity in agricultural soils. Journal
of Plant Nutrition and Soil Science, 169, 295–309.
Kögel-Knabner I, Amelung W, Cao Z, Fiedler S, Frenzel P, Jahn
R, Kalbitz K, Kölbl A, Schloter M. 2010. Biogeochemistry of
paddy soils. Geoderma, 157, 1–14.
Lefcheck J. 2016. PIECEWISESEM: Piecewise structural equation
modelling in R for ecology, evolution, and systematics.
Methods in Ecology and Evolution, 7, 573–579.
Lefcheck J S, Duffy J E. 2015. Multitrophic functional
diversity predicts ecosystem functioning in experimental
assemblages of estuarine consumers. Ecology, 96,
2973–2983.
1499
Li H, Wang J Q, Liu Q, Zhou Z F, Chen F L, Xiang D. 2019.
Effects of consecutive monoculture of sweet potato on soil
bacterial community as determined by pyrosequencing.
Journal of Basic Microbiology, 59, 181–191.
Li Q H, Wu F Z, Yang Y, Wang X Z. 2009. Effects of rotation and
interplanting on soil bacterial communities and cucumber
yield. Acta Agriculturae Scandinavica (Section B: Soil and
Plant Science), 59, 431–439.
Li Y, Chang S X, Tian L, Zhang Q. 2018. Conservation
agriculture practices increase soil microbial biomass carbon
and nitrogen in agricultural soils: A global meta-analysis.
Soil Biology and Biochemistry, 121, 50–58.
Liang B, Yang X, He X, Zhou J. 2011. Effects of 17-year
fertilization on soil microbial biomass C and N and soluble
organic C and N in loessial soil during maize growth. Biology
and Fertility of Soils, 47, 121–128.
Lithourgidis A S, Damalas C A, Gagianas A A. 2006. Long-
term yield patterns for continuous winter wheat cropping
in northern Greece. European Journal of Agronomy, 25,
208–214.
Liu C, Ding N, Fu Q, Brookes P C, Xu J, Guo B, Lin Y, Li H,
Li N. 2016. The influence of soil properties on the size
and structure of bacterial and fungal communities along
a paddy soil chronosequence. European Journal of Soil
Biology, 76, 9–18.
Liu E K, Yan C R, Mei X R, He W Q, Bing S H, Ding L P, Liu
Q, Liu S A, Fan T L. 2010. Long-term effect of chemical
fertilizer, straw, and manure on soil chemical and biological
properties in Northwest China. Geoderma, 158, 173–180.
Liu Z, Liu J, Yu, Z, Yao Q, Li Y, Liang A, Zhang W, Mi G, Jin
J, Liu X, Wang G. 2020. Long-term continuous cropping
of soybean is comparable to crop rotation in mediating
microbial abundance, diversity and community composition.
Soil & Tillage Research, 197, 104503.
Lu M, Yang Y, Luo Y, Fang C, Zhou X, Chen J, Yang X,
Li B. 2011. Responses of ecosystem nitrogen cycle to
nitrogen addition: A meta-analysis. New Phytologist, 189,
1040–1050.
Luo Y Q, Hui D F, Zhang D Q. 2006. Elevated CO2 stimulates net
accumulations of carbon and nitrogen in land ecosystems:
A meta-analysis. Ecology, 87, 53–63.
Manzoni S, Porporato A. 2009. Soil carbon and nitrogen
mineralization: Theory and models across scales. Soil
Biology & Biochemistry, 41, 1355–1379.
Marais A, Hardy M, Booyse M, Botha A. 2012. Effects of
monoculture, crop rotation, and soil moisture content on
selected soil physicochemical and microbial parameters
in wheat fields. Applied and Environmental Soil Science,
2012, 1–13.
Mcdaniel M D, Tiemann L K, Grandy A S. 2014. Does
agricultural crop diversity enhance soil microbial biomass
and organic matter dynamics? A meta-analysis. Ecological
Application, 24, 560–570.
Moore J M, Klose S, Tabatabai M A. 2000. Soil microbial
biomass carbon and nitrogen as affected by cropping
systems. Biology and Fertility of Soils, 31, 200–210.
1500 XING Ting-ting et al. Journal of Integrative Agriculture 2022, 21(5): 1488–1500
Nielsen U N, Ball B A. 2015. Impacts of altered precipitation
regimes on soil communities and biogeochemistry in arid
and semi-arid ecosystems. Global Change Biology, 21,
1407–1421.
Peralta A L, Sun Y, McDaniel M D, Lennon J T. 2018. Crop
rotational diversity increases disease suppressive capacity
of soil microbiomes. Ecosphere, 9, e02235.
Qiu S J, Gao H J, Zhu P, Hou Y P, Zhao S C, Rong X M, Zhang
Y P, He P, Christie P, Zhou W. 2016. Changes in soil carbon
and nitrogen pools in a Mollisol after long-term fallow or
application of chemical fertilizers, straw or manures. Soil &
Tillage Research, 163, 255–265.
R Core Team. 2018. R: A Language and Environment
for Statistical Computing. R Foundation for Statistical
Computing, Vienna.
Rengel Z. 2011. Soil pH, soil health and climate change.
In: Singh B P, Cowie A L, Chan K Y, eds., Soil Health
and Climate Change. Springer Berlin Heidelberg, Berlin,
Heidelberg. pp. 69–85.
Rosenberg M S, Adams D C, Gurevitch J. 2000. MetaWin:
Statistical Software for Meta-analysis. Version 2.0. Sinauer
Associates, Sunderland, Massachusetts.
Shahbazi B, Chelgani S C, Matin S S. 2017. Prediction of
froth flotation responses based on various conditioning
parameters by Random Forest method. Colloids and
Surfaces (A: Physicochemical and Engineering Aspects),
529, 936–941.
Shahid M, Nayak A K, Puree C, Tripathi R, Lal B, Gautam P,
Bhattacharyya P, Mohanty S, Kumar A, Panda B B, Kumar
U, Shukla A K. 2017. Carbon and nitrogen fractions and
stocks under 41 years of chemical and organic fertilization
in a sub-humid tropical rice soil. Soil & Tillage Research,
170, 136–146.
Shen Z, Penton C R, Lv N, Xue C, Yuan X, Ruan Y, Li R, Shen Q.
2018. Banana fusarium wilt disease incidence is influenced by
shifts of soil microbial communities under different monoculture
spans. Microbial Ecology, 75, 739–750.
Shipley B. 2009. Confirmatory path analysis in a generalized
multilevel context. Ecology, 90, 363–368.
Shipley B. 2013. The AIC model selection method applied to
path analytic models compared using a d-separation test.
Ecology, 94, 560–564.
Smith R G, Gross K L, Robertson G P. 2008. Effects of crop
diversity on agroecosystem function: Crop yield response.
Ecosystems, 11, 355–366.
Soil Survey Staff. 2006. Keys to Soil Taxonomy. Cornell
University, Ithaca, New York, USA.
Strobl C, Boulesteix A L, Zeileis A, Hothorn T. 2007. Bias in
random forest variable importance measures: Illustrations,
sources and a solution. BMC Bioinformatics, 25, 8–25.
Tang C, Rengel Z. 2003. Role of Plant Cation/Anion Uptake
Ratio in Soil Acidification. Marcel Dekker, New York. pp.
57–81.
Tian D S, Niu S L. 2015. A global analysis of soil acidification
caused by nitrogen addition. Environmental Research
Letters, 10, 024019.
Treseder K K. 2008. Nitrogen additions and microbial biomass:
A meta-analysis of ecosystem studies. Ecology Letters,
11, 1111–1120.
Zechmeister-Boltenstern S, Keiblinger K M, Mooshammer M,
Penuelas J, Richter A, Sardans J, Wanek W. 2015. The
application of ecological stoichiometry to plant-microbial-soil
organic matter transformations. Ecological Monographs,
85, 133–155.
Zhang H, Wu P, Yin A, Yang X, Zhang M, Gao C. 2017.
Prediction of soil organic carbon in an intensively managed
reclamation zone of eastern China: A comparison of multiple
linear regressions and the random forest model. Science
of the Total Environment, 592, 704–713.
Zhang J, Ai Z, Liang C, Wang G, Xue S. 2017. Response
of soil microbial communities and nitrogen thresholds of
Bothriochloa ischaemum to short-term nitrogen addition on
the Loess Plateau. Geoderma, 308, 112–119.
Zhang J, Qin J, Yao W, Bi L, Lai T, Yu X. 2009. Effect of
long-term application of manure and mineral fertilizers on
nitrogen mineralization and microbial biomass in paddy
soil during rice growth stages. Plant Soil and Environment,
55, 101–109.
Zhang Q, Miao F, Wang Z, Shen Y, Wang G. 2017. Effects
of long-term fertilization management practices on soil
microbial biomass in China’s cropland: A meta-analysis.
Agronomy Journal, 109, 1183.
Zhang X, Liu W, Zhang G, Jiang L, Han X. 2015. Mechanisms
of soil acidification reducing bacterial diversity. Soil Biology
and Biochemistry, 81, 275–281.
Zhong W, Cai Z, Yin L, He Z. 2007. Effects of the long-term
application of inorganic fertilizers on microbial community
diversity in rice-planting red soil as studied by using PCR-
DGGE. Acta Ecologica Sinica, 27, 4011–4018.
Zhou J, Shengxiu L I, Chen Z. 2002. Soil microbial biomass
nitrogen and its relationship to uptake of nitrogen by plants.
Pedosphere, 12, 251–256.
Zhou L Y, Zhou X H, Zhang B C, Lu M, Luo Y Q, Liu L L, Li
B. 2014. Different responses of soil respiration and its
components to nitrogen addition among biomes: A meta-
analysis. Global Change Biology, 20, 2332–2343.
Zhou Z, Wang C, Zheng M, Jiang L, Luo Y. 2017. Patterns and
mechanisms of responses by soil microbial communities
to nitrogen addition. Soil Biology and Biochemistry, 115,
433–441.
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