Environ Monit Assess (2014) 186:42234236

DOI 10.1007/s10661-014-3693-x

Soil respiration, labile carbon pools, and enzyme activities

as affected by tillage practices in a tropical ricemaizecowpea
cropping system
S. Neogi & P. Bhattacharyya & K. S. Roy & B. B. Panda &
A. K. Nayak & K. S. Rao & M. C. Manna

Received: 19 August 2013 / Accepted: 20 February 2014 / Published online: 8 March 2014
# Springer International Publishing Switzerland 2014

Abstract In order to identify the viable option of tillage

practices in ricemaizecowpea cropping system that
could cut down soil carbon dioxide (CO2) emission,
sustain grain yield, and maintain better soil quality in
tropical low land rice ecology soil respiration in terms of
CO2 emission, labile carbon (C) pools, water-stable
aggregate C fractions, and enzymatic activities were
investigated in a sandy clay loam soil. Soil respiration
is the major pathway of gaseous C efflux from terrestrial
systems and acts as an important index of ecosystem
functioning. The CO2C emissions were quantified in
between plants and rows throughout the year in rice
maizecowpea cropping sequence both under conventional tillage (CT) and minimum tillage (MT) practices
along with soil moisture and temperature. The CO2C
emissions, as a whole, were 24 % higher in between
plants than in rows, and were in the range of 23.478.1,
37.1128.1, and 28.6101.2 mg m2 h1 under CT and
10.760.3, 17.399.1, and 17.279.1 mg m2 h1 under
MT in rice, maize, and cowpea, respectively. The CO2
C emission was found highest under maize (44 %)
followed by rice (33 %) and cowpea (23 %) irrespective
of CT and MT practices. In CT system, the CO2C
S. Neogi : P. Bhattacharyya (*) : K. S. Roy : B. B. Panda :
A. K. Nayak : K. S. Rao
Division of Crop Production, Central Rice Research Institute,
Cuttack 753006 Orissa, India
e-mail: pratap162001@yahoo.co.in
M. C. Manna
Division of Soil Biology, Indian Institute of Soil Science,
Bhopal, India

emission increased significantly by 37.1 % with respect

to MT on cumulative annual basis including fallow. The
CO2C emission per unit yield was at par in rice and
cowpea signifying the beneficial effect of MT in maintaining soil quality and reduction of CO2 emission. The
microbial biomass C (MBC), readily mineralizable C
(RMC), water-soluble C (WSC), and permanganateoxidizable C (PMOC) were 19.4, 20.4, 39.5, and
15.1 % higher under MT than CT. The C contents in
soil aggregate fraction were significantly higher in MT
than CT. Soil enzymatic activities like, dehydrogenase,
fluorescein diacetate, and -glucosidase were significantly higher by 13.8, 15.4, and 27.4 % under MT
compared to CT. The soil labile C pools, enzymatic
activities, and heterotrophic microbial populations were
in the order of maize>cowpea>rice, irrespective of the
tillage treatments. Environmental sustainability point of
view, minimum tillage practices in ricemaizecowpea
cropping system in tropical low land soil could be
adopted to minimize CO2C emission, sustain yield,
and maintain soil health.
Keywords CO2C emission . Conventional tillage .
Minimum tillage . Cropping system . C fractions .
Enzymatic activity

Introduction
Carbon dioxide, a greenhouse gas responsible for enhanced global warming, has become an issue of major
concern due to its increased concentration in the

4224

atmosphere (IPCC 2007). It has been estimated that

agricultural practices contribute about 25 % of total
anthropogenic carbon dioxide (CO2) emissions and particularly soil CO2 emission is the important source of
gaseous carbon (C) flux in most ecosystems which
account for 6090 % of total ecosystem respiration
(Duxbury 1995). The relationship between production
and decomposition determines whether a system is a
sink or a source of atmospheric CO2. The CO2 emission
from soil to the atmosphere is the primary mechanism of
soil C loss (Parkin and Kasper 2003) and provides an
early indication of soil C level when changes in organic
carbon due to management practices are not detectable
within a short period (Bhattacharyya et al. 2013a). Soil
respiration includes root and microbial respiration, and
bulk turnover of organic matter which all contribute to
the release of CO2 (Hill et al. 2004). Thus, accurate
quantification of CO2 emissions from soil is important
for understanding of soil C flow and ecosystem resilience (Von Lutzow et al. 2006). The mechanism of soil
respiration is the key in assessing the global climate
change and carbon sequestration in soil. Actually, soil
CO2 emission integrates all components of soil CO2
production, including rhizospheric respiration and soil
microbial respiration. Hence, quantification of soil CO2
emission is very crucial for reducing C losses from soils
and to increase sequestration of atmospheric CO2 in soil.
Soil can act both as source and sink for atmospheric CO2
depending upon the management practices. Agricultural
operations and farm management practices can influence soil CO2 emission and C content in cropland
which, in turn, have effect on global warming due to
positive or negative feedback to C cycle. Soil CO2C
emission is increased by the land use change, biomass
burning, fossil fuel use, consumption of industrialized
products that demands high energy to manufacture, and
by tillage, that increases the biological activity and soil
C oxidation. Tillage practices can increase CO2 emission from soil by disrupting soil aggregates, incorporating plant residue and oxidizing soil organic carbon
(Barreto et al. 2009; Zhang et al. 2011). In the conventional tillage (CT), plowing followed by harrowing,
induces aggregate breakdown and increases C turnover
as a consequence of the exposure of organic matter to
microbes activity and their enzymes. Soil aggregation
has been considered the main mechanism of C protection of a wide range of soil classes (Fabrizzi et al. 2009).
Soil aggregation especially the macro-aggregates help in
protecting labile C and thus stabilize CO2C emission as

Environ Monit Assess (2014) 186:42234236

also reported by Manna et al. (2005). In addition, soil

tillage results in a higher contact between soil and residues and increases soil temperature, both processes that
favor organic matter decay and increase of soil CO2C
emission (Lal 2004). On the other hand, management
systems associated to minimum soil disturbances, such
as minimum tillage (MT), when combined with high
input of crop residues, favors soil C storage. The effect
of tillage systems on soil CO2C emission had been
investigated, and it was found that the emissions varied
depending upon the cropping systems and tillage practices (Sainju et al. 2005). Usually, short-term CO2C
emission after tillage is higher in the CT plots (La Scala
et al. 2006) or in some cases similar to the ones registered in the MT plots. On the other hand, higher emissions in MT than in CT systems have also been observed, which indicates that the effect of soil tillage on
soil CO2C emission could be related to the duration,
season, soil type, cropping system, soil moisture and
temperature, the mechanism of soil C stabilization, and
C stock. Cropping system can influence CO2 emission
by affecting the quantity and quality of residue returned
to soil (Al-kaisi and Yin 2005). Tillage and cropping
sequence have significant impact on soil microbial activity and the decomposition processes that transform
the plant-derived C to soil organic matter (SOM) and
CO2 emission by changing the soil environment viz. soil
moisture, soil temperature, soil labile C pools, soil biological activities (soil enzymatic activities and microbial
population), soil pH, and soil aeration (Amos et al.
2005). Therefore, the effect of MT and CT practices in
ricemaizecowpea cropping systems on the quantification of CO2 emissions as well as soil labile C pools
and associated biological activities is needed for C
budgeting and modeling of gaseous C emission. In light
of that, the present study was conducted to (1) quantify
the annual CO2 emission and (2) establish relationship
between CO2 emissions with soil labile C pools and
biological activities from ricemaizecowpea cropping
system under minimum tilled and conventionally tilled
soils.

Materials and methods

Experimental site
The study site is situated at the experimental farm of the
Central Rice Research Institute Cuttack (202657.84

Environ Monit Assess (2014) 186:42234236

4225

N, 85563.41E; 24 m above mean sea level) in the

eastern part of India. The climate is tropical, with short
winter and lengthy hot summer period and heavy cyclonic rainfall during monsoon season. The average
annual temperature of the region was 28 C which was
one of the criteria of tropical climate as per the Koppen
climate classification (Diaz and Eischeid 2007) was
concerned. The mean annual precipitation is around
1,500 mm. The characteristics of the initial soil properties at the study site are presented in tabular form
(Table 1).
Crop establishment and treatments
This study was conducted in CT and MT treatments in
continuous rice (cv. Naveen; spacing, plant row,
15 cm20 cm), maize (cv. Super 36; spacing, plant
row, 20 cm30 cm), and cowpea (cv. PDM 54; spacing,
plantrow, 15 cm20 cm) cropping system for 3 years.
In this cropping system, the rice was sown on 12 June
2010 and was harvested on 17 October 2010. The maize
crop was sown in the field on 16 November 2010 and
was harvested on 27 February 2011. On the other hand,
cowpea was grown for short duration (sowing on 11
March 2011 and harvested on 22 May 2011). The tillage
treatments were imposed in a complete randomized
block design with three replications. In CT treatment,
the field was plowed and puddled thoroughly to 20-cm
depth and leveled before the wet direct seeding.
Moldboard plow was used before the cultivation of rice
crop during monsoon, and chisel plow was used during

Table 1 Characteristics of initial soil properties at the study site

Soil parameters

Concentration

Clay (%)

25.9

Silt (%)

21.6

Sand (%)

52.5

Bulk density (Mg m3)

1.41

pH (H2O)

6.06

Cation exchange capacity (c mol (p+) kg1)

15.2

Electrical conductivity (dS m1)

0.5

Total C (%)

0.49

Total N (%)

0.05

Olsens extractable P (mg kg1)

14.2

Available K (mg kg1)

356

other seasons. Water was given to the field through

surface irrigation without runoff return system whenever it was necessary. Primary tillage treatments involving
moldboard plowing were applied in the wet season of
2010 (June 2010) and secondary tillage operations involving chisel plowing were carried out in the months of
November 2010 and March 2011 just before planting of
maize and cowpea, respectively. Under MT (disked
once, cultivated once up to 10 cm depth, and residue
removed), rice was dry direct seeded during June 2010
with manual dibbling method causing minimum soil
disturbance. In a similar way, maize and cowpea were
sown during the months of November 2010 and March
2011, respectively, by dibbling method.
Soil sampling and storage
Sampling was done during the various growth stages of
rice (maximum tillering, T; panicle initiation, PI; grain
filling, GF; and maturity, M), maize (fifth leaf initiation,
LI; silking, SK; cobbling, CB; and maturity, M), and
cowpea (third trifoliate leaf initiation, LIA; flowering,
FL; pod forming, PF; and maturity, M) from three
replicated plots (10 m10 m) of each tillage treatments
distributed in randomized block design. Five soil cores
(8-cm diameter) samples from each replicated plots
were taken at 020 cm depth for rice, maize, and cowpea, and composite mixture were made. A portion of
each field-moist composite soil sample was kept at 4 C
for microbial biomass, soil enzymes analysis, and microbial population estimation, while the other portion
was air-dried in shade. Three hundred grams of air dried
undisturbed sample were used for soil aggregate analysis and the rest was passed through a 2-mm sieve prior to
soil labile C fractions analysis. Soil temperature and
moisture was determined by time domain refractometer
(TDR; TRIME-EZC, M/S Imko Micromoduletechnik
Gmbh, Germany).
Carbon dioxide emission measurement
Infrared gas analyzer (IRGA)-based (model EGM-4, PP
system, Haverhill, MA, USA) field measurement technique was used for quantifying CO2 emission rates in
enclosed chamber over a specified time. The CO2 emission was measured with an environmental gas monitor
chamber (closed dynamic chamber) attached to a data
logger. A flag was placed as a marker in the plot where
CO2 emission was measured throughout the study

4226

period in order to produce representable datasets and

eliminate sampling error and biasness of sampling
(Bhattacharyya et al. 2012a, b). The chamber was
15 cm tall, 10 cm in diameter, and had capacity to
measure CO 2 emission from 0 to 9.99 g CO 2
C m2 h1. The chamber was placed at the soil surface
in maize and cowpea and on base plate (3 cm height) in
rice for 2 min for each plot until CO2 emission measurement was recorded in the data logger. The CO2 emission
was recorded from the plots in the plantplant (PP) (in
between plant) and rowrow (RR) (in between rows)
position for rice, maize, and cowpea, and also from plots
of bare soil (without any tillage practice and crops). All
the multipoints measurements (to reduce site-specific
variability) were taken between 9:00 to 11:00 hours in
the morning and between 15:00 to 17:00 hours in the
afternoon which was considered as the representative
time of sampling in this region to reduce diurnal variability of CO2 flux (Bhattacharyya et al. 2012a, b,
2013a). Moreover, the measurements were done at
higher frequency (34 days intervals) throughout the
year to get a clear picture of annual CO2 fluxes from
the system by the infrared gas analyzer having higher
precision. However, this method only considered the
soil CO 2 C flux (i.e., soil respiration) but not
encompassed the total ecosystem C flux (i.e., net ecosystem exchange or ecosystem respiration) which could
be measured by eddy covariance system (Bhattacharyya
et al. 2013a, b). The average between the morning and
evening emissions was considered as the daily mean
emission. The observations were taken on the Julian
days of 198, 201, 204, 207, 210, 213, 216, 219, 222,
225, 228, 231, 234, 237, 240, 243, 246, 249, 252, 255,
258, 261, 264, 267, 270, 273, 276, 279, 282, 285, 288,
291, 296, 301, 307, 311, 315, 322, 326, 332, 334, 337,
340, 344, 348, 351, 355, 359, 362, and 365 of the year
2010 and on 3, 7, 10, 13, 16, 20, 24, 27, 31, 34, 37, 40,
44, 47, 50, 53, 56, 59, 62, 65, 68, 71, 74, 77, 81, 87, 90,
95, 100, 105, 110, 113, 116, 120, 123, 126, 130,
134, 138, 141, 145, 148, 151, 154, 158, 162, 166,
171, 175, 179, 183, 186, 190, 193, and 197 of the
year 2011.
Soil labile C parameters analysis
Soil microbial biomass-C (MBC) was measured by
modified chloroform fumigationextraction method
with fumigation at atmospheric pressure (Witt et al.
2000). Readily mineralizable C (RMC) content of the

Environ Monit Assess (2014) 186:42234236

soil samples was estimated after extraction with 0.5 M

K2SO4 (Inubushi et al. 1991) followed by wet digestion
of the soil extract with dichromate (Vance et al. 1987).
The water-soluble carbohydrate C (WSC) was estimated
followed by the procedure of Haynes and Swift (1990).
Permanganate oxidizable carbon (PMOC) was determined by following the method described by Blair
et al. (1995) with a few modifications. Dry soil (3 g)
was weighed into 50 ml centrifuge tubes, and 30 ml of
20-mM KMnO4 was added. The centrifuge tubes were
shaken for 15 min and centrifuged for 5 min at
2,000 rpm. The absorbance of the supernatant and standards was read at 565 nm. The change in the concentration of KMnO4 was used to estimate the amount of
carbon oxidized, assuming that 1 mM KMnO4 is consumed in the oxidation of 0.75 mM or 9 g of carbon.
Total C (TC) was estimated by dry combustion in CHN
analyzer (FLASH 2000 Organic Elemental analyzer,
M/s Thermo Scientific).

Aggregate size fractions

Soil samples (020 cm depth) were collected for aggregate size fractions analysis after the harvest of the cowpea crop of ricemaizecowpea cropping system. The
undisturbed soil samples (>2 mm) were used for aggregate separation by wet sieving method (Camberdella
and Elliott 1992). Two 100-g subsamples were slaked,
capillary rewetted, and wet sieved by double-stage
Yodders apparatus through a series of three sieves to
obtain four size fractions: >2, 2502, 53250, and
<53 m. Aggregate fractions retained on each sieve
were transferred into a container, dried at 65 C in an
oven, weighted, and grinded to pass 150-m size before
C analysis.

Soil enzymatic activities and microbial populations

Dehydrogenase activity was determined by reduction of
2,3,5-triphenyltetrazolium chloride (TTC) (Casida et al.
1964). Fluorescein diacetate (FDA) hydrolysis activity
measurements were made following the method of
Adam and Duncan (2001). The -glucosidase activity
was assayed following the procedure of Eivazi and
Tabatabai (1988). The heterotrophic microbial populations were cultured by using the media of Rand
et al. (1975).

Environ Monit Assess (2014) 186:42234236

4227

Table 2 Effect of conventional tillage (CT) and minimum tillage

(MT) practices on crop yield, cumulative CO2 emission
Crop
type

Treatment

Rice
Maize
Cowpea

Yield
(t ha1)

Cumulative
CO2C
emission
(kg ha1)

CO2C emission
per unit yield
(kg CO2C
per kg yield)

CT

6.0b

1,716.5b

0.29a

MT

5.1a

1,290.2a

0.25a

CT

8.5b

2,677.0b

0.31b

MT

7.7a

1,858.6a

0.24a

CT

0.24b

1,280.2b

5.33a

MT

0.18a

1,038.2a

5.76a

In each column, values (mean of three replicated observations) of

the treatments followed by different letters are significantly
(p<0.05) different by Duncans multiple range test (DMRT) for
each crop type

multiple range test (DMRT) at the 0.05 level of probability using statistical software SPSS 20.0 (Statistical
Package for Social Sciences).

Results and discussions

Yield of crops
There was significant (p<0.05) increase in the crop
yield under CT treatment in comparison to that of under
MT treatment irrespective of the crop type. The grain
yield varied 5.16, 7.78.5, and 0.180.24 t ha1 in rice,
maize, and cowpea, respectively (Table 2). In rice and
cowpea, emission per unit yield basis was at per under
MT and CT, whereas in maize, it was significantly
higher under CT than MT (Table 2).

Statistical analysis

Emission of CO2C

Individual character datasets were subjected to analysis

of variance, and means were separated by Duncan

There were distinct temporal variations of CO2C emission noticed under MT and CT practices in different

Soil CO2-C flux

(mg m-2 h-1)

200.0

180.0

CT system

Bare soil

P-P soil

R-R soil

Maize

160.0
140.0

Cowpea

120.0

Rice

100.0
80.0
60.0
40.0
20.0
0.0

Soil CO2-C flux

(mg m-2 h-1)

198 207 216 225 231 243 252 261 270 279 288 301 315 332 340 351 362 7

200.0
180.0
160.0
140.0
120.0
100.0
80.0
60.0
40.0
20.0
0.0

16 27 37 47 56 65 74 87 100 113 123 134 145 158 162 179 190

MT system
Maize
Cowpea
Rice

198 207 216 225 234 243 252 261 270 279 288 301 315 332 340 351 362 7

Year 2010

Fig. 1 Soil carbon dioxide (CO2C) flux from bare soil, plant to
plant (PP), and row to row (RR) portion under conventional
tillage (CT) (a) and minimum tillage (MT) (b) practices in a rice

16 27 37 47 56 65 74 87 100 113 123 134 145 154 166 179 190

Julian days of year

Year 2011

maizecowpea cropping system. Bars and half bars indicate the

standard deviations from the means of three replicated observations (p<0.05)

4228

Environ Monit Assess (2014) 186:42234236

Table 3 Effect of conventional tillage (CT) and minimum tillage (MT) practice on soil labile C pools at different crop growth stages under
ricemaizecowpea cropping system
Soil C pools

Treatment Rice
T

MBC (g g1)

CT

RMC (g g )
WSC (g g1)

PI

77.2a

MT
1

Maize
GF

LI

Cowpea
SK

CB

LIA

FL

PF

95.5a

86.5a 53.1a 183.9a 219.4a 205.4a 126.9a 169.8a 209.9a 191.0a 107.7a

103.1b 121.5b

97.1b 69.4b 237.8b 272.3b 229.1b 139.7b 206.1b 243.5b 213.5b 129.7b

CT

28.5a

38.5a

36.8a 13.4a

74.0a

91.6a

86.9a

49.8a

49.9a

63.7a

58.3a

28.8a

MT

39.4b

49.5b

46.0b 17.4b

91.5b 104.4b

93.7b

58.4b

70.8b

84.2b

67.5b

32.7b

CT

12.6a

26.3a

22.6a 11.2a

38.2a

55.8a

51.2a

24.0a

22.5a

36.3a

30.8a

11.4a

MT

23.5b

39.1b

33.5b 12.5a

55.7b

68.6b

59.8b

32.6b

43.4b

56.8b

40.1b

15.3a

KMnO4C (g g1) CT

120.9a 149.7a 142.0a 81.4a 344.2a 426.1a 404.3a 231.7a 224.2a 277.5a 263.3a 150.9a

MT

149.4b 170.5b 153.1b 95.3b 425.5b 485.4b 435.9b 271.6b 277.1b 316.1b 283.8b 176.8b

In each column, values (mean of three replicated observations) of the treatments followed by different letters are significantly (p<0.05)
different by Duncans multiple range test (DMRT) in each growth stage under each crop for each labile C parameter. The growth stages
included were maximum tillering (T), panicle initiation (PI), grain filling (GF) and maturity (M) in rice, the fifth initiation (LI), silking (SK),
cobbling (CB) and maturity (M) in maize, the third trifoliate leaf formation (LIA), flowering (FL), pod forming (PF), and maturity (M) in
case of cowpea
MBC microbial biomass C, RMC readily mineralizable C, WSC water-soluble carbohydrate C, and KMnO4C permanganate oxidizable C

crops (Fig. 1a, b). Significantly (p<0.05) higher emissions of CO2C were noticed under CT in comparison to
that of MT practice (Fig. 1a, b). The CO2C emissions
were significantly higher in PP in comparison to that of
RR soil (Fig. 1a, b) irrespective of the MT and CT
practices. The emission of CO2C was in the order of
maize>cowpea>rice irrespective of the MT and CT
practices (Fig. 1a, b).
The PP, CO2C emission varied in between 22.1
75.6 and 30.189.4 mg m2 h1 in different stages of
rice growth under MT and CT practices, respectively,
and the highest emissions were observed in the PI stage
(258th Julian days in 2010).
In maize, the PP, CO2C emission varied in between 35.8125.3 and 48.8146.2 mg m2 h1 under
MT and CT practices, respectively, and the highest

emissions were observed at the SK stage of the crop

development (61st Julian days in 2011).
In cowpea, the maximum PP, CO2C emission was
noticed during the period of LIA to FL (92nd Julian
days in 2011) stage.
The CO2C emissions during the fallow periods of
the year also varied significantly under MT and CT
practices with a range of 16.156.5 and 17.193.2,
respectively, while in bare soil the emission varied in
between 3.1 and 9.5 mg m2 h1. The cumulative CO2
C emission varied from 1,290.2 to 1,716.5, 1,858.6 to
2,677.0, and 1,038.2 to 1,280.2 kg ha1 in MT and CT
practices under rice, maize, cowpea, respectively
(Table 2).
Soil CO2 efflux was often significantly influenced by
tillage practices and crop species (Sainju et al. 2005). It

Table 4 Effect of conventional tillage (CT) and minimum tillage (MT) practices on mass percentage and total C content of different soil
aggregate size classes
Total C (g kg1)

Mass percentage (%)

Treatment

>2 mm

250 m2 mm

53250 m

<53 m

>2 mm

250 m2 mm

53250 m

<53 m

CT

8.8a

49.2a

39.5b

2.5b

8.7a

4.6a

4.1a

5.2a

MT

11.2b

52.1b

33.4a

3.3a

10.5b

4.9b

4.8b

6.0b

In each column, values (mean of three replicated observations) followed by different letters are significantly (p<0.05) different by Duncans
multiple range test (DMRT)

The growth stages included were maximum tillering (T), panicle initiation (PI), grain filling (GF) and maturity (M) in rice, the fifth initiation (LI), silking (SK), cobbling (CB) and maturity
(M) in maize, the third trifoliate leaf formation (LIA), flowering (FL), pod forming (PF), and maturity (M) in case of cowpea. In each column, values (mean of three replicated observations)
of the treatments followed by different letters are significantly (p<0.05) different by Duncans multiple range test (DMRT) in each growth stage under each crop for each enzyme

4.6b

3.7a
7.6a

8.3b
8.7b

8.1a
6.2a

8.0b
5.3b

4.4a
8.3a

9.0b
9.5b

8.7a
7.0a

8.8b
3.8b

2.9a
6.8a

7.5b
7.9b
7.3b

7.2a
5.4a

MT

17.1b
30.3b
35.9b
29.1b
23.6b
41.9b
50.9b
40.3b
12.3b
21.9b
25.9b
21.1b
MT

CT
soil)
Heterotrophic population (log CFU g

3.5b

2.9a

173.2b

12.6a
26.8a
29.3a
20.0a
17.4a
37.0a
40.4a
27.6a
9.1a
19.3a
21.2a
14.5a
CT
-Glucosidase activity (g p-nitrophenol g1 h1)

5.5b
6.2b
5.4b
6.1b
9.8b
10.9b
9.6b
3.1b
5.0b
4.9b
MT

5.5b

5.1a

277.9b
309.5b

5.4a
4.4a

271.3b
316.4b

5.2a
9.1a

507.8b
565.5b

9.6a
7.7a

503.2b
133.4b

2.6a
4.6a

214.1b
238.5b

4.9a
3.9a

209.0b
MT

CT
FDA activity (g fluorescein g1 h1)

147.7a
257.8a
271.7a
219.5a
269.9a
471.1a
496.4a
443.2a
113.8a
198.6a
209.3a
169.1a
CT
Dehydrogenase activity (g TPF g1day1)

M
PF
FL
LIA
LI
PI

GF

M
T

SK

CB

Cowpea
Maize
Rice
Treatment
Enzymatic parameters and
microbial population

is evident from the study that CT increased CO2 emission compared with MT at corresponding measurement
dates irrespective of crop types, and that the emission of
CO2 was more in PP in comparison to RR. The CO2
C emission was significantly (p<0.05) higher in CT
than MT, and was consistent with the reports of AlKaisi and Yin (2005). This was attributed to rapid physical release of CO2 (entrapped CO2 from soil pore
spaces and aggregates released due to pulverization
caused by tillage) trapped in the soil air space due to
soil physical disturbance (Jackson et al. 2003). The
higher emission of CO2C in CT was due to greater soil
volume that was disturbed under CT relative to MT
(Reicosky et al. 2005). Tillage can result in an immediate short-term outburst of CO2 due to a decrease in
partial pressure of CO2 in soil air, followed by disturbance in soil aggregation and pores, and sudden release
of CO2 from the soil solution (Reicosky et al. 2005).
Under raindrop and irrigation impact, the soil surface
may have sealed in the CT treatment, temporarily trapping CO2. The raindrop and irrigation led to increase the
moisture content of soil which promoted higher rate of
microbial activity and respiration in soil as also reported
by Bhattacharyya et al. (2012b). The trapped CO2 may
have released, when the water drained off and soil
surface becomes dried. In other way, CT practices accompanied by rainfall and irrigation could have increased soil microbial activities, thereby increasing C
mineralization and CO2 emission. The difference in
CO2 emission with crop sequence irrespective of the
CT or MT treatment suggests that the type of crop may
have influenced root and microbial respiration. In this
study, the CO2C emission was highest under maize
followed by cowpea and rice. The CO2C emission
during the different crops varied due to the difference
in water management practices and root system. It is
well studied that 48 % of C loss as CO2 emission
during the crop-growing season was contributed by
respiration from plant roots and rhizosphere (Iqbal
et al. 2009), provided that root respiration during
the off-season was negligible. In this study, maize
having very proliferated, extensive root system and
increased biomass in comparison to that of the rice
and cowpea (data not presented), contributed the
maximum emission. The submergence anaerobic
condition in rice (35 cm water depth) prevents
the CO2 emission as compared to the maize and
cowpea which were cultivated under aerobic
condition.

4229
Table 5 Effect of conventional tillage (CT) and minimum tillage (MT) practice on soil enzymatic activities and microbial population at different crop growth stages under ricemaize
cowpea cropping system

Environ Monit Assess (2014) 186:42234236

4230

Environ Monit Assess (2014) 186:42234236

In this study, the highest CO2 flux was observed

under maize in the CT system. Conventional tillage
accelerates organic C oxidation to CO2 by improving
soil aeration, increasing contact between soil and crop
residues and exposing aggregate-protected organic matter to microbial attack (Iqbal et al. 2009). Soil labile C,
like MBC, has been included in the current soil monitoring concept due to its rapid response and high sensitivity to management practices and environmental
changes. Soil organic carbon (SOC) and MBC acts as
important binding agents for aggregation (Six et al.
2000, 2002). The CT generally breakdowns

Soil labile C pools

The labile C fractions like MBC (53.1272.3 g g1),
RMC (13.4104.4 g g1), WSC (11.268.6 g g1),
and PMOC (81.4485.4 g g1) were all increased
significantly (p<0.05) under MT with respect to that of
CT in ricemaizecowpea cropping system (Table 3).
The maximum labile pools were observed in maize
followed by cowpea and rice irrespective of the tillage
practices (Table 3). The highest quantities of soil labile
C pools were found in PI, SK, and FL stages of rice,
maize, and cowpea, respectively (Table 3).

Rice
P-P

100.0

y = 12.567x + 65.296
R = 0.5455

Rice
R-R

100.0

80.0

80.0
60.0

60.0
40.0

40.0

20.0

20.0

0.0

Soil CO2-C flux (mg m-2 h-1)

-3.0

-2.0

-1.0

Maize
P-P

200.0

160.0

-3.0

-2.0

y = 10.126x + 48.841
R = 0.5251

0.0
0.0

1.0

2.0

y = 28.687x + 107.49
R = 0.7533

-2.0

0.0

Maize
R-R

200.0

160.0

120.0

120.0

80.0

80.0

40.0

40.0

0.0
-1.0 0.0

Cowpea
P-P

150.0

120.0

2.0

3.0

y = 18.531x + 85.9
R = 0.7237

-2

-1

Cowpea
R-R

150.0

120.0

y = 12.242x + 79.314
R = 0.8559

90.0

60.0

60.0

30.0

30.0

0.0
-2.0

4.0

y = 26.62x + 90.637
R = 0.7552

0.0
1.0

90.0

-4.0

2.0

0.0
0.0

2.0

4.0

-4.0

-2.0

0.0

2.0

4.0

Change in soil temperature (C)

Fig. 2 Correlation (p<0.05) between soil CO2C flux with the

change in soil temperature in rice plant to plant (PP) (a), rice row
to row (RR) (b), maize plant to plant (PP) (c), maize row to row

(RR) (d), cowpea plant to plant (PP) (e), and cowpea row to row
(RR) (f) system under conventional tillage (CT) practice

Environ Monit Assess (2014) 186:42234236

4231

et al. 2010). The greater CO2 flux in maize system than

in other crop was probably related to increased root and
microbial respiration due to higher root and rhizosphere
growth that may have increased substrate availability for
soil microorganisms (Sainju et al. 2005).

mechanically macro-aggregates (Borie et al. 2006) and

decreases the content of soil labile C and MBC
(Curaqueo et al. 2011). MT has positive effects on
aggregates and their binding agents, greater MBC, and
turnover of higher SOC stock (Helgason et al. 2010;
Curaqueo et al. 2011; Zhang et al. 2012). The physically
protected C in soil macro- or micro-aggregates under
CT system was very prone to microbial attack, as the
capacity of C stabilization is much less in CT system as
compared to that of under MT practices. Hence, the CT
practice leads to higher amount gaseous C flux and low
reserve of the TC. The soluble C fraction is an important
part with respect to SOM turnover as it acts as a readily
decomposable substrate for soil microorganisms and as
a short-term reservoir of plant nutrients (Garcla-orenes

Rice
P-P

There were significant (p<0.05) differences in soil aggregates and labile C fractions observed under the CT
and MT practices. The amount of soil macro-aggregates
(>2 mm and 250 m2 mm soil aggregate class size)
increased significantly under MT in comparison to that
of under CT (Table 4). The mass percentage of microaggregate (<53 m soil aggregate class size) was also

y = 11.397x + 52.748
R = 0.5734

100.0

Soil aggregate size classes and associated carbon

Rice
R-R

100.0

80.0

y = 9.4504x + 36.923
R = 0.5389

80.0

60.0

60.0

40.0

40.0

20.0

20.0

0.0

Soil CO2-C flux (mg m-2 h-1)

-3.0

-2.0

0.0

1.0

0.0

2.0

-3

200.0

Maize
P-P

y = 31.5x + 86.277
R = 0.7374

160.0

-2.0

-1.0

-1.5

-1.0

-2

-1

Maize
R-R

200

160

120.0

120

80.0

80

40.0

40

0.0
-0.5 0.0

Cowpea
P-P

y = 13.509x + 53.289
R = 0.7771

0
0.5

1.0

-2

1.5

150.0

e
120.0

-1

Cowpea
R-R

150.0

120.0

y = 18.185x + 60.852
R = 0.7809

y = 14.902x + 75.701
R = 0.7672

90.0

90.0
60.0

60.0

30.0

30.0

0.0

0.0
-3.0

-2.0

-1.0

0.0

1.0

2.0

-4.0

-2.0

0.0

2.0

4.0

Change in soil temperature

Fig. 3 Correlation (p<0.05) between soil CO2C flux with the

change in soil temperature in rice plant to plant (PP) (a), rice row
to row (RR) (b), maize plant to plant (PP) (c), maize row to row

(RR) (d), cowpea plant to plant (PP) (e), cowpea row to row (R
R), and (f) system under minimum tillage (MT) practice

4232

Environ Monit Assess (2014) 186:42234236

increased significantly under MT (3.3 %) in comparison

to that of CT (2.5 %; Table 4). The TC content in each
aggregate soil class size increased significantly under
MT treatment in comparison to that of under the CT
(Table 4).
Soil aggregation is the result of the rearrangement of
particles, flocculation, and the cementation of microstructures and micro-aggregates (Duiker et al. 2003), a
process which depends on the contents of TC, biota,
ionic bonds, clay, and carbonates (Bronick and Lal
2005). The accumulation of soil organic matter (SOM)
due to physical protection occurs because of the influence of aggregation in the formation of fracture planes
(here referring to the sides of aggregates connected by
cementing agents) that act as physical barriers to
100

y = 1.6424x - 8.6914
R = 0.607

Rice
P-P

90
80

90

80

Rice
R-R

70

70

y = 1.8052x - 28.112
R = 0.5986

60

60

50

50

40

40

30

30

20

20

10

10

0
30

35

c
Soil CO2-C flux (mg m-2 h-1)

microbial action. In MT system, the aggregates and the

soil structure are maintained due to the lack of plowing,
which, over time, results in an increase of macroaggregates (Barreto et al. 2009; Fabrizzi et al. 2009)
and an accumulation of TC (Paustian et al. 2000). This
increase in the formation of large macro-aggregates may
have a positive impact on the development of crops in
years with water limitations because greater intraaggregate porosity contributes to the storage of water
over longer periods of time. This effect provides benefits
for the physical properties of the soil by leading to
greater infiltration of water, a decrease in erosion
(Franzluebbers 2002), and a greater physical protection
of the SOM (Balabane and Plante 2004) which in turn
resist the microbial C decomposition and generate

200.0

40

45

50

Maize
P-P

55

30

60

y = 5.3562x - 14.448

R = 0.7199

200.0

35

40

45

Maize
R-R

50

55

60

y = 5.8796x - 37.407

R = 0.7595

160.0

160.0
120.0

120.0

80.0

80.0

40.0

40.0

0.0
10.0

e
150.0

15.0

20.0

25.0

30.0

Cowpea y = 3.6206x + 30.166

R = 0.7712
P-P

0.0
10.0

150.0

15.0

20.0

Cowpea
R-R

25.0

30.0

y = 3.2587x + 24.535
R = 0.7318

120.0

120.0

90.0

90.0
60.0

60.0

30.0

30.0
0.0

0.0
5.0

10.0

15.0

20.0

25.0

5.0

10.0

15.0

20.0

25.0

Soil volumetric moisture (v/v) %

Fig. 4 Correlation (p<0.05) between soil CO2C flux with the

soil volumetric moisture content (%) in rice plant to plant (PP)
(a), rice row to row (RR) (b), maize plant to plant (PP) (c),

maize row to row (RR) (d), cowpea plant to plant (PP) (e),
cowpea row to row (RR), and f system under conventional tillage
(CT) practice

Environ Monit Assess (2014) 186:42234236

4233

(50.9 gp-nitrophenol g1 h1) activity was obtained

in the maize under the MT (Table 5). The highest
heterotrophic microbial population (9.5 log CFU) was
also observed in maize under MT (Table 5). The enzymatic activities and microbial population varied significantly in each growth stages under different crop types.
The maximum activity was found in PI, SK, and FL
stages of rice, maize, and cowpea, respectively
(Table 5).
The activity of assayed enzymes was generally well
correlated with the labile C content. Dehydrogenase
activity basically depends on the metabolic state of the
soil biota. Total microbial activity in terms of fluorescein
diacetate has been used to determine the amount of
active microflora-producing extracellular enzymes
(Adam and Duncan 2001). In this study, a significant

lowered CO2C emission as compared to that of under

CT system. The TC found in micro-aggregates is considered to be more resistant to microbial decomposition,
leading to an accumulation of SOM for a long period
(Six et al. 2002). Micro-aggregates have been considered a key factor in the formation of new macroaggregates (Six et al. 2000) due to the protection of
intra-aggregates TC.
Soil enzymatic activities
Soil enzyme activities and microbial populations were
strongly influenced by the tillage practices and type of
the crops (Table 5). In the present study, the maximum
dehydrogenase (565.5 g TPF g 1 day 1 ), FDA
(10.9 g fluorescein g1 h1), and -glucosidase

80

y = 1.6344x - 24.18
R = 0.6032

Rice
P-P

y = 1.7065x - 43.55
R = 0.6025

Rice
R-R

60

70

50

60
50

40

40

30

30

20

20

10

10

0
30

Soil CO2-C flux (mg m-2 h-1)

70

200.0

35

40

45

50

55

60

30

y = 4.7138x - 21.449
R = 0.7123

Maize
P-P

d
200.0

160.0

160.0

120.0

120.0

80.0

80.0

40.0

40.0

0.0
10.0

15.0

20.0

Cowpea
P-P

25.0

30.0

0.0
10.0

120.0

y = 3.0926x + 15.45 150.0

R = 0.7199
120.0

90.0

90.0

60.0

60.0

30.0

30.0

150.0

35

0.0

40

45

Maize
R-R

50

55

60

y = 4.252x - 30.337
R = 0.7115

15.0

20.0

25.0

30.0

Cowpea
R-R
y = 3.2194x + 0.6009
R = 0.7582

0.0
5.0

10.0

15.0

20.0

25.0

5.0

10.0

15.0

20.0

25.0

30.0

Soil volumetric moisture (v/v) %

Fig. 5 Correlation (p<0.05) between soil CO2C flux with the

soil volumetric moisture content (%) in rice plant to plant (PP)
(a), rice row to row (RR) (b), maize plant to plant (PP) (c),

maize row to row (RR) (d), cowpea plant to plant (PP) (e),
cowpea row to row (RR), and (f) system under minimum tillage
(MT) practice

4234

increase in dehydrogenase and FDA activity occurred

under the MT. Minimum tillage has redistributed soil
carbon and microbial biomass towards the surface soil,
increased potential enzyme activities, and reduced emissions of carbon dioxide (Drijber et al. 2000).

Environ Monit Assess (2014) 186:42234236

dependent and becomes moisture dependent when

soil dried up (Smith et al. 2003). Similar observations were noticed in this study, where maize and
cowpea were grown under aerobic condition as
compared to rice which was cultivated under submergence condition.

Relationship between CO2C emission with soil

temperature and moisture
Regression analysis of CO2C emission with change of
soil temperature at 020 cm at the time of CO2 measurement revealed that CO2C emission was linearly
related with change in soil temperature under CT (R2 =
0.52 to 0.85; Fig. 2af) and MT practices (R2 =0.53 to
0.78; Fig. 3af). Changes in soil temperature were more
under CT than MT. The CO2C emission was linearly
related with volumetric moisture content (in percent)
both under CT practices (R2 =0.59 to 0.77; Fig. 4af)
and MT practices (R2 =0.60 to 0.75; Fig. 5af) throughout the cropping system.
Increased soil water content and decreased in change
in soil temperature in MT compared with CT as evident
from this study have also been reported by Al-Kaisi and
Yin (2005). High CO2 emission usually occurs when
soil temperature is higher and soil water content and
substrate C availability are adequate (Bajracharya et al.
2000). The study showed that CO2 emission was well
related with change in soil temperature and volumetric
moisture content of soil irrespective of the crops and
tillage practices. The change in soil temperature was
taken into account to nullify the seasonal variation of
soil temperature throughout the year. Less regression
coefficient between CO2 emission and change in soil
temperature were noticed under rice because of water
stagnation in rice which act as an insulator to soil
temperature change. Similarly, the low regression coefficients were observed with CO2 emission and moisture
content in rice as it was cultivated in submerged condition. Management practices, such as tillage, cropping
systems, and irrigation significantly altered CO2 emissions even at the same soil temperature and moisture
content (Smith et al. 2003). The results from this study
also suggested that tillage practices partly modified
soil temperature and moisture content, which, in
turn, influenced microbial activities, C mineralization, and soil CO2 emissions. The soil temperature
is the primary factor governing annual CO2 emission (Bhattacharyya et al. 2013a). The release of
CO2 by aerobic respiration is primarily temperature

Conclusion
Interaction effect of tillage and crops influenced the soil
labile C content and enzymatic and soil biological activities which in turn affected soil respiration, i.e., CO2
emission in a tropical ricemaizecowpea cropping system. The PP soil CO2 emission was more strongly
correlated with soil temperature and moisture than that
of RR. Soil aggregate C is well protected in MT than in
CT; as a result, the CO2 emission was less in MT. In
tropical rice-based cropping system, annual CO2 emissions from croplands can be minimized by adopting MT.
From a sustainable environment point of view, minimum tillage practices in ricemaizecowpea cropping
system was an adoptable intervention compared to conventional tillage in respect to CO2C emission, sustain
yield, and soil health. This approach of quantification of
soil respiration in relation to tillage practices could be
extended to ricerice, ricemaize, ricegreen gram, and
riceblack gram cropping system in similar ecologies.
However, long-term studies are needed to determine the
interaction effects of tillage, water management, and
cropping sequence on CO2C emission and soil C sequestration under various rice-based cropping systems
in tropical region.
Acknowledgments The work has been partially supported by
the grant of ICAR-NAIP, Component-4 (2031), Soil organic
carbon dynamics vis--vis anticipatory climatic changes and crop
adaptation strategies, NICRA and CRRI. Part of the findings is
the Ph.D. work of Mr. S. Neogi. The valuable guidance of Dr. D.C.
Uprety, Dr. V.R. Rao, Dr. S.N. Singh, Dr. Sudhir Kochhar, and Dr.
T.K. Adhya is acknowledged. Technical support was provided by
the technical staff of the division of Crop Production CRRI.

References
Adam, G., & Duncan, H. (2001). Development of a sensitive and
rapid method for the measurement of total microbial activity
using fluorescein diacetate (FDA) in a range of soils. Soil Biol
Biochem, 33, 943951.

Environ Monit Assess (2014) 186:42234236

Al-Kaisi, M. M., & Yin, X. (2005). Tillage and crop residue effects
on soil carbon and carbon dioxide emission in corn-soybean
rotation. J Environ Qual, 34, 437445.
Amos, B., Arkebauer, T. J., & Doran, J. W. (2005). Soil surface
fluxes of greenhouse gases in an irrigated maize-based
agroecosystem. Soil Sci Soc Am J, 69, 387395.
Bajracharya, R. M., Lal, R., & Kimble, J. M. (2000). Diurnal and
seasonal CO2C flux from soil as related to erosion phases in
central Ohio. Soil Sci Soc Am J, 64, 286293.
Balabane, M., & Plante, A. F. (2004). Aggregation and carbon
storage in silty soil using physical fractionation techniques.
Eur J Soil Sci, 55, 415427.
Barreto, R. C., Madari, B. E., Maddock, J. E. L., Machado, P. L. O.
A., Torres, E., Franchini, J., & Costa, A. R. (2009). The impact
of soil management on aggregation, carbon stabilization and
carbon loss as CO2 in the surface layer of a Rhodic Ferralsol in
Southern Brazil. Agric Ecosyst Environ, 132, 243251.
Bhattacharyya, P., Nayak, A. K., Mohanty, S., Tripathi, R., Shahid,
M., Kumar, A., Raja, R., Panda, B. B., Roy, K. S., Neogi, S.,
Dash, P. K., Shukla, A. K., & Rao, K. S. (2013a). Greenhouse
gas emission in relation to labile soil C, N pools and functional microbial diversity as influenced by 39 years long-term
fertilizer management in tropical rice. Soil Tillage Res, 129,
93105.
Bhattacharyya, P., Neogi, S., Roy, K. S., Dash, P. K., Tripathi, R.,
& Rao, K. S. (2013b). Net ecosystem CO2 exchange and
carbon cycling in tropical lowland flooded rice ecosystem.
Nutr Cycl Agroecosyst, 95, 133144.
Bhattacharyya, P., Roy, K. S., Neogi, S., Adhya, T. K., Rao, K. S.,
& Manna, M. C. (2012a). Effects of rice straw and nitrogen
fertilization on greenhouse gas emissions and carbon storage
in tropical flooded soil planted with rice. Soil Tillage Res,
124, 119130.
Bhattacharyya, P., Roy, K. S., Neogi, S., Chakravorti, S. P.,
Behera, K. S., Das, K. M., Bardhan, S., & Rao, K. S.
(2012b). Effect of long term application of organic amendment on C storage in relation to global warming potential and
biological activities in tropical flooded soil planted to rice.
Nutr Cycl Agroecosyst, 94, 273285.
Blair, G. J., Lefroy, R. D. B., & Lisle, L. (1995). Soil carbon
fractions based on their degree of oxidation, and the development of a carbon management index for agricultural systems. Aust J Agric Res, 46, 14591466.
Borie, F., Rubio, R., Rouanet, J. L., Morales, A., Borie, G., &
Rojas, C. (2006). Effects of tillage systems on soil characteristics, glomalin and mycorrhizal propagules in a Chilean
Ultisol. Soil Tillage Res, 88, 253261.
Bronick, C. J., & Lal, R. (2005). Soil structure and management: a
review. Geoderma, 124, 322.
Camberdella, C. A., & Elliott, E. T. (1992). Particulate organic
matter changes across a grassland cultivation sequence. Soil
Sci Soc Am J, 56, 777783.
Casida, L. E., Klein, D. A., & Santoro, T. (1964). Soil dehydrogenase activity. Soil Sci, 98, 371376.
Curaqueo, G., Miguel, B. J., Acevedo, E., Rubio, R., Cornejo, P.,
& Borie, F. (2011). Effects of different tillage system on
arbuscular mycorrhizal fungal propagules and physical properties in a Mediterranean agroecosystem in central Chile. Soil
Tillage Res, 113, 1118.
Diaz, H. F., & Eischeid, J. K. (2007). Disappearing alpine tundra
Kppen climatic type in the western United States.

4235
Geophysical Research Letter, 34, L18707. doi:10.1029/
2007GL031253.
Drijber, R. A., Doran, J. W., Parkhurst, A. M., & Lyon, D. J.
(2000). Changes in soil microbial community structure with
tillage under long-term wheat-fallow management. Soil Biol
Biochem, 32, 14191430.
Duiker, S. W., Rhoton, F. E., Torrent, J., Smeck, N. E., & Lal, R.
(2003). Iron (hydr)oxide crystallinity effects on soil aggregation. Soil Sci Soc Am J, 67, 606611.
Duxbury, JM (1995) The significance of agricultural greenhouse
gas emissions from soil of tropical agroecosystems. In: R. Lal
(ed.). Soil management and greenhouse effect. Lewis: Boca
Raton, FL. pp. 279291
Eivazi, F., & Tabatabai, M. A. (1988). Glucosidases and galactosidases in soils. Soil Biol Biochem, 20, 601606.
Fabrizzi, K. P., Rice, C. W., Amado, T. J. C., Fiorin, J.,
Barbagelata, P., & Melchiori, R. (2009). Protection of soil
organic C and N in temperate and tropical soils: effect of
native and agroecosystems. Biogeochemistry, 92, 129143.
Franzluebbers, A. J. (2002). Soil organic matter stratification ratio
as an indicator of soil quality. Soil Tillage Res, 66, 95106.
Garcla-orenes, F., Guerrero, C., Roldan, A., Mataix-Solera, J.,
Cerda, A., Campoy, M., Zornoza, R., Barcenas, G., &
Caravaca, F. (2010). Soil microbial biomass and activity under
different agricultural management systems in a semiarid
Mediterranean agroecosystem. Soil Tillage Res, 109, 110115.
Haynes, R. J., & Swift, R. S. (1990). Stability of soil aggregates in
relation to organic constituents and soil water content. J Soil
Sci, 41, 7383.
Helgason, B. L., Walley, F. L., & Germida, J. J. (2010). No-till soil
management increases microbial biomass and alters community profiles in soil aggregates. Appl Soil Ecol, 46, 390397.
Hill, P. W., Marshall, C., Harmens, H., Jones, D. L., & Farrar, J.
(2004). Carbon sequestration: do N inputs and elevated atmospheric CO2 alter soil solution chemistry and respiratory C
losses? Water Air Soil Pollut, 4, 177186.
Inubushi, K., Brookes, P. C., & Jenkinson, D. S. (1991). Soil
microbial biomass C, N and ninhydrin-N in aerobic and
anaerobic soils measured by fumigation-extraction method.
Soil Biol Biochem, 23, 737741.
IPCC. (2007). Climate Change 2007: the physical science basis,
contribution of Working Group-I to the fourth assessment
report of the Intergovernmental Panel on Climate Change.
Cambridge: Cambridge University Press.
Iqbal, J., Hu, R., Lin, S., Hatano, R., Feng, M., Lu, L., Ahamadou,
B., & Du, L. (2009). CO2 emission in a subtropical red paddy
soil (Ultisol) as affected by straw and N fertilizer applications: a case study in Southern China. Agric Ecosyst Environ,
131, 292302.
Jackson, L., Calderson, F. J., Scow, K. L., Steenwerth, K. M., &
Rolston, D. E. (2003). Responses of soil microbial processes
and community structure to tillage events and implications
for soil quality. Geoderma, 114, 305317.
La Scala, N., Bolonhezi, D., & Pereira, G. T. (2006). Short-term
soil CO2 emission after conventional and reduced tillage of a
no-till sugarcane area in southern Brazil. Soil Tillage Res, 96,
244248.
Lal, R. (2004). Soil carbon sequestration impacts on global climate
change and food security. Science, 304, 16231627.
Manna, M. C., Swarup, A., Wanjari, R. H., Ravankar, H. N.,
Mishra, B., & Saha, M. N. (2005). Long-term effect of

4236
fertilizer and manure application on soil organic carbon storage, soil quality and yield sustainability under sub-humid and
semi-arid tropical India. Field Crop Res, 93, 264280.
Parkin, T. B., & Kasper, T. C. (2003). Temperature controls on
diurnal carbon dioxide flux: implication for estimating soil
carbon loss. Soil Sci Soc Am J, 67, 17631772.
Paustian, K., Six, J., Elliott, E. T., & Hunt, H. W. (2000).
Management options for reducing CO2 emissions from agricultural soils. Biogeochemistry, 48, 147163.
Rand, M. C., Greenberg, A. E., Taras, M. J., & Franson, M. A.
(1975). Standard methods for the examination of water and
waste water. Washington: American Public Health Association.
Reicosky, D. C., Lindstrom, M. J., Schumacher, T. E., Lobb, D. E.,
& Malo, D. D. (2005). Tillage-induced CO2 loss across and
eroded landscape. Soil Tillage Res, 81, 183194.
Sainju, U. M., Whitehead, W. F., & Singh, B. P. (2005). Biculture
legume-cereal cover crops for enhanced biomass yield and
carbon and nitrogen. Agron J, 97, 14031412.
Six, J., Elliott, E. T., & Paustian, K. (2000). Soil macroaggregate
turnover and microaggregate formation: a mechanism for C
sequestration under no-tillage agriculture. Soil Biol Biochem,
32, 20992103.
Six, J., Feller, C., Denef, K., Ogle, S. M., Sa, J. C. D., & Albrecht, A.
(2002). Soil organic matter, biota and aggregation in temperate
and tropical soilseffects of no-tillage. Agronomie, 22, 755775.

Environ Monit Assess (2014) 186:42234236

Smith, K. A., Ball, T., Conen, F., Dobbie, K. E., Massheder, J., &
Rey, A. (2003). Exchange of greenhouse gases between soil
and atmosphere: interactions of soil physical factors and
biological processes. Eur J Soil Sci, 54, 779791.
Vance, E. D., Brookes, P. C., & Jenkinson, D. S. (1987). An
extraction method for measuring soil microbial biomass carbon. Soil Biol Biochem, 19, 703707.
Von Lutzow, M., Kogel-Knabner, I., Ekschmitt, K., Matzner, E.,
Guggenberger, G., Marschner, B., & Flessa, H. (2006).
Stabilization of organic matter in temperate soils: mechanisms and their relevance under different soil conditionsa
review. Eur J Soil Sci, 57, 426445.
Witt, C., Gaunt, J. L., Galicia, C. C., Ottow, J. C. G., & Neue, H. U.
(2000). A rapid chloroform fumigationextraction method
for measuring soil microbial biomass carbon and nitrogen
in flooded rice soils. Biol Fertil Soils, 30, 510519.
Zhang, H., Wang, X., Feng, Z., Pang, J., Lu, F., Ouyang, Z.,
Zheng, H., Liu, W., & Hui, D. (2011). Soil temperature and
moisture sensitivities of soil CO2 efflux before and after
tillage in a wheat field of Loess Plateau, China. J Environ
Sci, 23, 7986.
Zhang, S., Li, Q., Zhang, X., Wei, K., Chen, L., & Liang, W.
(2012). Effects of conservation tillage on soil aggregation
and aggregate binding agents in black soil of Northeast
China. Soil Tillage Res, 124, 196202.