Catena 166 (2018) 171–180

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Catena
journal homepage: www.elsevier.com/locate/catena

Soil organic carbon pool changes in relation to slope position and land-use in T
Indian lower Himalayas
⁎
Pritpal Singh , Dinesh K. Benbi
Department of Soil Science, Punjab Agricultural University, Ludhiana, India

A R T I C LE I N FO A B S T R A C T

Keywords: The increase in the atmospheric abundance of CO2 has raised interest in the potential of soils to sequester carbon
C sequestration (C), which inter alia depends on antecedent C level. While inventories of soil organic carbon (SOC) have been
Soil erosion developed for different ecosystems at field or global scales, studies at hill-slope level where SOC redistributes by
Labile C erosion remain scarce. In addition, quantitative information on the variation in SOC stocks and organic matter
Recalcitrant C
quality within hill-slopes would allow to make more realistic estimations of the C sequestration potential of
Grasslands
Croplands
landscapes while suggesting improved land management. The main objective of this study was to evaluate the
Land-use impact of slope position and land-use (grassland versus cropland) on SOC stocks and organic matter quality
Macro-aggregates (proportion of aggregate associated C and labile pools; water extractable, WEOC; hot water soluble, HWC; po-
tassium permanganate oxidizable, KMnO4-C; microbial biomass, MBC). A total of 108 soil samples from the
0–0.15 m layer were collected from 4 slope positions (forested hilltops, middle slope with bare soils, backslopes
under grasslands, cropped bottomlands) at nine catenas with 3 replicates per position. The greatest SOC stocks
were found at hilltops and backslopes with respectively 10.1 ± 0.64 and 10.1 ± 0.57 Mg C ha−1. SOC stocks
decreased by 17% to 8.4 ± 0.65 Mg C ha−1 in bottomlands and by 58% to 4.2 ± 0.26 Mg C ha−1 at backslopes.
The soil organic matter from the hilltops showed the greatest proportion of recalcitrant components (62%)
followed by the backslope and bottomland positions with ~42%, and middle slope position with 9%. In addition
to showing a greater occurrence of labile C fractions, bare and eroded backslope soils were also depleted in water
soluble C (WEOC and HWC), KMnO4-C and microbial biomass C. The results showed that there is considerable
potential for C sequestration at middle and backslope position through land rehabilitation by either planting
trees or vegetation by grass.

1. Introduction
The atmospheric concentration of carbon dioxide (CO2) has in-
creased globally by 47% from 278 parts per million (ppm) in the pre-
industrial era to 409 ppm in 2017 (NOAA, 2017). During 2002–2011,
the atmospheric concentration of CO2 increased at a rate
(2 ppm year−1) higher than any previous decade since direct mea-
surements of atmospheric concentration commenced (Ciais et al.,
2013). Increasing concentration of CO2 is forcing global climate
change; and efforts are being made to stabilize the atmospheric abun-
dance of CO2. A number of strategies have been advocated and are
being implemented to achieve this goal, which inter alia include C se-
questration in soil and vegetation (Nieder and Benbi, 2008). The rate
and magnitude of C sequestration in soil besides depending on soil
properties, climatic conditions, land-use and management is influenced
to a great extent by antecedent C level. In the last three decades, several
studies have enumerated the effect of agricultural management on
organic matter turnover and soil's feed-back to global climate change. A
number of management practices including intensification of agri-
culture, no-till farming, planting of cover crops, improved nutrient
management and crop residue recycling have been recommended for
enhanced soil C sequestration (Lal, 2004; Powlson et al., 2011); and
estimates have been made for C sequestration potential from these ac-
tivities (Follett et al., 2000; Smith, 2004). However, the realizable po-
tential for C sequestration in soils may be much less than that estimated
because of the effect of existing soil organic C (SOC) level and the soil's
capacity to store C. Therefore, an assessment of SOC content and stocks
is imperative for evaluation of C sequestration potential of soils (Batjes,
1996; Eswaran et al., 1993). While several studies have investigated the
distribution of soil carbon and SOC stock changes in different ecosys-
tems at a range of scales such as field scale (Paustian et al., 1992; Smith
et al., 1997; Schelsinger and Lichter, 2001), regional, sub-national and
national scales (Arrouays et al., 2001; Bonfatti et al., 2016; Jones et al.,
2005; Milne et al., 2007), studies at hill-slope level remain scarce. This

⁎
Corresponding author at: Krishi Vigyan Kendra, Khokhar Khurd, Mansa, Punjab 151 501, India.
E-mail address: jasppsingh@gmail.com (P. Singh).

https://doi.org/10.1016/j.catena.2018.04.006
Received 11 July 2017; Received in revised form 28 March 2018; Accepted 4 April 2018
Available online 10 April 2018
0341-8162/ © 2018 Elsevier B.V. All rights reserved.
P. Singh, D.K. Benbi
Fig. 1. Soil sampling sites and schematic representation of sampling plan at typical hill-slopes in the Siwalik foothills of Punjab, India.
could lead to large uncertainties in estimates of regional SOC stocks.
Studies at the field scale use site-specific data in relatively homogenous
conditions and have limited wider applicability. Studies at large scale
such as Chaplot et al. (2010) in Laos showed that SOC stocks are sig-
nificantly affected by land-use, rainfall and hill-slope level. On sloping
landscapes, considerable amounts of SOC, especially black carbon are
lost by water erosion (Chaplot et al., 2005). Landscape level differences
in soil C in relation to land cover and topography in the eastern US
showed that effects of topography were usually secondary to those of
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Catena 166 (2018) 171–180
land cover (Garten and Ashwood, 2002).
Spatial distribution of SOC vary widely across landscapes leading to
large uncertainties in the SOC budget especially for mountainous
landscapes susceptible to erosion where a tight coupling between geo-
morphic processes and soil C turnover exists (Doetterl et al., 2012).
Combining anisotropy analysis with discrete wavelet transform, Guo
et al. (2018) observed that topography and local climate may have a
strong influence in controlling SOM spatial distribution in mountain
regions. Organic C storage and loss from soil depends on soil type (Six
P. Singh, D.K. Benbi
et al., 2002; Worrall et al., 2012), topography such as inclination, slope
length etc. (Chaplot and Poesen, 2012), vegetation cover (Valentin
et al., 2008) and agricultural practices (Benbi et al., 2012). Studies
relating changes in SOC to soil erosion, deposition pattern and topo-
graphic position (Johnson et al., 2000; Moorman et al., 2004; Ritchie
et al., 2007) suggest that information on soil redistribution and topo-
graphic position could be used to understand SOC dynamics of the
landscapes. Topographic position, soil erosion and land-use not only
alter the quantity of SOC, but also its composition. Quantification of
SOC stocks and pool distribution along geomorphic gradients showed
that size and composition of the SOC pools were strongly related to
geomorphic position (Doetterl et al., 2012) and the loss of SOC was
related to its lability or decomposability (Murty et al., 2002). Labile C
pools are reported to respond differentially to land-use and manage-
ment (Ashagrie et al., 2007; Golchin and Asgari, 2008). The composi-
tion of eroded C changes from the local to the catchment level, because
the erosion and the sedimentation processes fractionate SOC (Chaplot
et al., 2005). Aggregates are important agents of SOC storage and its
protection against decomposition (Six et al., 2002). Extent of C reten-
tion in soil depends on soil aggregation (Benbi et al., 2016; Carter,
1996); short term C storage occurs in macro-aggregates (> 0.25 mm)
and long-term C storage in micro-aggregates (< 0.25 mm). The
knowledge on the relative abundance of labile C pools and distribution
of water stable aggregates (WSA) and their C density in relation to
topographic position and land-cover can help understand the effect of
erosion processes on SOC dynamics and potential for soil C sequestra-
tion in hill-slope soils.
Despite several studies on global SOC inventories, or at research site
on experiments with similar soil or landscape conditions, quantitative
data on SOC stocks and organic matter quality along typical hill-slopes,
which are globally lacking, are absolutely necessary to mitigate climate
change through appropriate management of soils. There is a crucial
need for field data on soil C distribution along catena and on the impact
of the key factors of climate, hydrology, parent material, soil fertility,
biological activity, vegetation patterns and land cover – all of which
control the level of organic matter in soils (Jenny, 1941). This will
allow for improving strategies for land management. Such studies are
also expected to provide requisite inputs for regional or global simu-
lation models intended to assess the possible impacts of the changes
expected in land-cover and climate on SOC in relation to potential
greenhouse gas emissions. If it is hypothesized that greater C seques-
tration can be achieved by soil and land-cover management, then it is
important to significantly improve our understanding of how differ-
ences in topographic position and land-use interact to affect soil C
storage along typical hill-slopes. But such information along elevation
catena, which integrates the effect of topography, land-use and de-
gradation, and vegetation cover is particularly lacking for sub-tropical
and tropical regions in Asia. Our objectives were to study variations in
soil C and organic matter quality along typical hill-slopes of lower Hi-
malayas so as to understand the potential impacts of slope position and
land-use.
2. Materials and methods
2.1. Site description
The study area is situated in a sub-mountain (Siwalik foothills) region
of north Indian state of Punjab and comprises gently to moderately
sloping, undulating and dissected land (Fig. 1). These ranges are com-
posed of unconsolidated sediments brought down by rivers from the
main Himalayan ranges located further north. The valleys are covered
with thick gravel and alluvium. The region represents the most fragile
ecosystem where soil erosion is a serious problem. Over the years
natural vegetation has diminished; grasslands have been converted to
cropland and forest products removed to meet food, fuel and other
requirements of the growing population (Pandit et al., 2007). All of
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Catena 166 (2018) 171–180
these have contributed towards accelerated soil erosion. Typically, an
elevation catena is interspersed with hilltops at the highest elevation
followed by eroded sites down the slope and grasslands and croplands
at the lowest elevation. The climate of the region is classified as semi-
arid sub-tropical with strong monsoonal influence. May is the hottest
month with mean monthly maximum and minimum temperatures of
38.2 °C and 21.9 °C, respectively (Kaur and Hundal, 2008a). January is
the coldest month, with mean monthly maximum temperature of
19.2 °C and mean minimum temperature of 5.2 °C. Average annual
rainfall averages 1122 mm, of which ~86% is received during summer
(June–October; maize growth season) and the remaining 14% during
winter season (November–April; wheat growth season) (Kaur and
Hundal, 2008b). The soil moisture regimes of the study sites were Udic
and Ustic.
2.2. Sampling locations
Nine different sampling locations with similar physiographic con-
ditions were selected. The locations were selected in such a way that
each represented an elevation catena involving forested hilltops, middle
slopes with bare soils followed by backslopes under grassland and
cropped bottomland. Hilltop soils (elevation
398–422 m above mean sea level, amsl) had minimum disturbance since
decades, and were under natural vegetation of mixed shrubs and un-
managed volunteer tall bushy grasses. The middle slopes (elevation
281–356 m amsl), which had been losing top soil layer had no vegeta-
tion. Backslope grassland soils (elevation 251–269 m amsl) were under
natural dwarf grass species. The cropped bottomlands (elevation
254–272 m amsl) had been under maize (Zea mays L.) -wheat (Triticum
aestivum L.) cropping for the last over 20 years. Maize and wheat were
grown in a sequence with maize during summer and wheat during
winter. On the sampled sites, maize crop was fertilized with
115–125 kg N and 25–35 kg P2O5 ha−1. In addition, farm yard manure
(FYM) at 10 Mg ha−1 was applied before seeding maize, once in two
years. Wheat was fertilized at 120 kg N and 60 kg P2O5 ha−1. At some
locations, where soils were deficient in potassium, an application of
30 kg K2O ha−1 was made to wheat Nitrogen, phosphorus and po-
tassium were applied through urea (46% N), diammonium phosphate
(18% N and 46% P2O5) and muriate of potash (60% K2O), respectively.
Both the crops were adequately irrigated with groundwater.
2.3. Soil sampling and sample preparation
Surface soil samples (0–0.15 m depth) were collected from hilltops,
middle slopes, and backslope grasslands and croplands at the bottom at
9 selected locations. A schematic representation of the sampling plan
along the elevation catena at every site is shown in Fig. 1. Soil samples
were collected in May–June 2013 after the harvest of wheat in crop-
lands. At each site 3 pseudo-replications were established and a total of
108 soil samples (9 location × 4 slope positions × 3 replications) were
collected. Soil samples were collected with a metallic soil core sampler
(inner diameter 7 cm). Top 0.15 m depth was chosen as it represented
the soil layer that experienced the greatest disturbance due to tillage
and erosion and received differential input of exogenous and plant-
mediated C. One portion of the sample was passed through a 2 mm
sieve for analysis of soil chemical and biological properties and the
other portion was retained without sieving for aggregate analysis.
2.4. Sample analyses
2.4.1. Physical analyses
Soil bulk density (Db) was determined by core method. Soil cores,
collected from each sampling site, were oven dried at 105 °C for 24 h,
and dry soil weight was recorded (Blake and Hartge, 1986). The Db
(Mg m−3) was calculated as:
P. Singh, D.K. Benbi
Ws
Db =
Vt
where, ‘Ws’ is weight of soil (Mg) and Vt is the volume of soil sample
(m3).
For aggregate analysis, soil clods (> 8.0 mm size, passed through
10 mm sieve and retained on 8.0 mm sieve) were spread uniformly on a
nest of sieves (2.0, 1.0, 0.50, 0.25, 0.11 and 0.053 mm) and wet-sieved
to allow for aggregate size separation (Yoder, 1936). The soil samples
on the sieves were slaked for 20 min in water and thereafter, nest of
sieves was oscillated up and down by a pulley arrangement for 30 min
at a frequency of 30 cycles/min in salt-free water. Water stable ag-
gregates (WSA) of different sizes were collected from the respective
sieves separately and weighed after oven drying at 50 °C. Water stable
aggregates were classified into two main aggregate size classes namely
macro-aggregates (> 2.0–0.25 mm) and micro-aggregates
(> 0.053–0.25 mm). Macro-aggregates were further classified into
coarse macro-aggregates (CMacA, > 2.0 mm) and meso-aggregates
(MesoA, < 2.0–0.25 mm). Similarly, micro-aggregates were classified
into coarse micro-aggregates (CMicA, < 0.25–0.11 mm) and fine micro-
aggregates (FMicA, < 0.11–0.053 mm). The mean weight diameter
(MWD) of aggregates was calculated as:
n
MWD (mm) = ∑i =1 Xi × Wi
where, n is the number of aggregate size fractions, Xi is the mean dia-
meter (mm) of the sieve size class and Wi is the weight of aggregates (g)
retained on ith sieve (van Bevel, 1950). The aggregate ratio was com-
puted as:
[%Water stable macroaggregates > 0.25 mm]
Aggregate ratio =
[%Water stable microaggregates < 0.25 mm]
Aggregates of different sizes were analyzed for total organic C (TOC)
by wet digestion method (Snyder and Trofymow, 1984). Carbon pre-
servation capacity (CPC) of each size class of aggregates was calculated
by using C concentration (%) and proportion of aggregates (g kg−1) in
the ith class by the following relationship.
(Aggregate associated C )i × (Water stable aggregate)i
CPC (g kg−1 soil) =
100
2.4.2. Chemical analyses
The sieved soil samples were analyzed for pH (1:2 soil: water sus-
pension) with a glass electrode, electrical conductivity (EC, 1:2 soil:
water supernatant) using a conductivity meter, and particle size dis-
tribution by pipette method (USDA, 1930).
Total organic carbon in whole soil and aggregates was determined
by reacting with 1 N K2Cr2O7 solution at 150 °C for 1 h (Snyder and
Trofymow, 1984). The time and temperature of heating were standar-
dized in a preliminary experiment conducted with a number of soils
from the experimental region. Heating time of 1 h at 150 °C yielded
results similar to those obtained with dry combustion method. Total
organic C stock (Mg C ha−1) in the 0.15 m soil layer was computed by
multiplying TOC concentration (%) with measured soil bulk density
(Mg m−3) and depth (m) of sampling using the following relationship:
TOC (Mg C ha −1) = TOC (%) × Soil depth (m) × Bulk density (Mg m−3)
× 100
Change in TOC stocks in soils at hilltops, middle slopes and crop-
lands were computed with reference to grassland soils.
Water-extractable organic C (WEOC) was determined by shaking
10 g soil with 20 ml deionized water for 1 h (Benbi et al., 2014). Hot
water soluble carbon (HWC) was determined by moderately boiling
20 g soil with 100 ml distilled water for 1 h under reflux condenser. The
amount of C in the extract was determined by chromosulfuric acid
method (Schulz et al., 2003). Potassium permanganate oxidizable C
(KMnO4-oxidizable C) was determined by oxidation with 33 mM
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Catena 166 (2018) 171–180
KMnO4 solution for 24 h (Blair et al., 1995). The amount of non-oxi-
dizable C, hence after referred as non-labile C, was determined as the
difference between TOC and KMnO4-oxidizable C. Information on
KMnO4-oxidizable C was used to a calculate carbon management index
(CMI) as per the procedure described by Blair et al. (1995). The KMnO4-
C for grassland soils was taken as the reference.
CMI = CPI × LI × 100
where, CPI is the C pool index and LI is the lability index. The CPI and
the LI were calculated as:
TOC(hilltop / eroded / cropland) (g kg−1 soil)
CPI =
TOC(grassland) (g kg−1 soil)
L(hilltop / eroded / cropland)
LI =
L(grassland)
where, L refers to the C lability, calculated as
Content of labile C (KMnO4 oxidizable C ) (g kg−1 soil)
L=
Content of non lable C (g kg−1 soil)
Different fractions of TOC under a gradient of oxidizing conditions
were determined using sulfuric acid (H2SO4)–aqueous solution ratios of
0.5:1, 1:1 and 2:1, which correspond to 12 N, 18 N and 24 N H2SO4,
respectively (Chan et al., 2001). The concentration of organic C de-
termined using the three acid-aqueous solution ratios allowed separa-
tion of TOC into the following four fractions of decreasing oxidiz-
ability/lability.
CVL (very labile C) = organic C oxidizable under 12 N H2SO4
CL (labile C) = 18 N–12 N H2SO4 oxidizable C
CLL (less labile C) = 24 N–18 N H2SO4 oxidizable C
CR (recalcitrant C) = TOC–24 N H2SO4 oxidizable C.
Carbon mineralization (Cmin) was studied for 45 days in the la-
boratory by conducting aerobic incubation at 25 °C temperature and
field capacity moisture as per the procedure described by Benbi et al.
(2014). Fifty grams of each air-dried soil was wetted to field capacity
moisture (50% water filled pore space) and placed in a conical flask
along with vials containing 10 ml 1 M NaOH to trap evolved carbon
dioxide (CO2). Alkali traps were replaced daily during the first 13 days
and every alternate day thereafter. Evolved CO2 was determined by
titrating alkali in the traps with 0.1 M HCl using phenolphthalein as an
indicator. Carbon mineralization rate during 43rd and 45th days of
incubation was taken as basal soil respiration (BSR). Microbial biomass
C (MBC) was determined at the end of incubation by Chloroform Fu-
migation Extraction (CFE) method (Vance et al., 1987) using a recovery
factor (KEC) of 0.41 (Voroney and Paul, 1984). Mineralization quotient
(qM) was calculated as the ratio of Cmin to TOC (μg Cmin μg TOC−1)
(von Lützow et al., 2002).
Respiratory quotient (qCO2) was computed as the ratio of BSR to
microbial biomass C (μg CO2 basal-C h−1 × μg biomass C−1) × 103
(Anderson and Domsch, 1986). The microbial quotient (qmic) was
calculated as the ratio of MBC to TOC (μg biomass C μg TOC−1)
(Anderson and Domsch, 1989).
2.5. Statistical analyses
Experimental data were statistically analyzed by one-way analysis
of variance (ANOVA) using SPSS for Windows 16.0 (SPSS Inc., Chicago,
U.S.A.). Mean separation for different land-uses and eroded soils was
performed using Duncan's multiple range test (DMRT) and were tested
for significance at 95% confidence interval (p < 0.05). To identify
organic C pools and other soil properties that may contribute towards
group separation of topographic positions, discriminant function ana-
lysis (DFA) was performed. The results of DFA have been graphically
presented in two dimensions for the first two discriminant functions
P. Singh, D.K. Benbi Catena 166 (2018) 171–180

Table 1 slope soils. This showed that with reference to the non-eroding upslope
Particle size distribution and other properties of surface soils (0–0.15 m) at soils, the middle slopes had lost the greatest amount of C
forested hilltops, middle slope eroded soils, backslope grasslands and croplands (5.9 Mg C ha−1) followed by croplands at the bottom of the slope
in Siwalik foothill region of northern India. Values in parenthesis indicate (1.7 Mg C ha−1). Higher TOC stocks in grassland and hilltop soils may
standard error from mean.
be attributed to greater plant-mediated C input through root biomass,
Property Hilltop Eroded Grassland Cropland root exudates and rhizodeposition and little or no soil disturbance. The
a a a
soils under grassland could have also been enriched with deposition of
pH 7.78 7.60 7.54 (0.113) 7.32a (0.037)
C from the eroding slopes. Ritchie et al. (2007) also reported that up-
(0.09) (0.06)
E.C. (dS m−1) 0.321b 0.271a 0.253a 0.234a land eroding soils had significantly less organic C than soils in deposi-
(0.018) (0.02) (0.013) (0.007) tion areas. Equivalent stocks of TOC in soils at the hilltops and under
Sand (%) 68.1a 68.2a 61.9a (1.07) 64.6a (0.95) the grasslands show that despite the differences in topographic position
(1.02) (1.23)
and type of vegetation- shrubs and tall grasses at hilltops and dwarf
Silt (%) 19.9a 20.8a 20.9a (1.36) 20.3a (1.00)
(0.53) (1.16)
grasses in grasslands, these could maintain comparable C input-output
Clay (%) 12.0a 11.0a 17.2b (1.23) 15.0b (1.07) balance. Under relatively undisturbed conditions, as was prevalent at
(1.08) (0.91) hilltops, there is a near-equilibrium between C input and C output
Bulk density 1.57b 1.57b 1.49a (0.011) 1.50a (0.005) fluxes resulting in maintenance of SOC status (Benbi et al., 2014),
(Mg m−3) (0.023) (0.021)
whereas cultivation of undisturbed soils results in loss of organic C as
Mean values for a soil property followed by different letter differ significantly was evident from 16.8% lower TOC stocks in cropland soils compared
(p < 0.05) by Duncan's multiple range test (DMRT). to grassland and hilltop soils. It has been reported that increased C
residence due to reduced disturbance in the absence of tillage in the
explaining maximum variance, which are generally considered im- grasslands results in higher C storage, compared with croplands
portant. (Conant et al., 2001; Soussana et al., 2004). A number of studies have
shown that organic C stocks in soils under cropland are lower (by 11 to
70%) compared to soils under grassland (Gerzabek et al., 2005; Jiao
3. Results and discussion
et al., 2009; Xun et al., 2010). David et al. (2009) reported 30–50%
reduction in TOC pool due to conversion of prairies to cultivation.
3.1. Basic soil properties
Smaller C loss under cropland in our study may be attributed to the
exogenous input of organic C through regular application of FYM and
The soils at all the slope positions were non-saline (EC
deposition of C from eroding slopes. Several studies have shown that
0.27–0.32 dS m−1), near neutral to slightly alkaline in reaction
though TOC storage in cropland soils was improved with fertilizer and
(pH 7.3–7.8) and had sand and silt contents ranging between
manure application, but it was still lower than soils under grassland
61.9–68.2% and 19.9–20.9%, respectively (Table 1). Soils at lower
(Blair et al., 2006; Gami et al., 2009). Lower C stocks in cropland soils
slope positions, under grassland and cropland, had significantly
compared to grasslands suggest that there is considerable opportunity
(p < 0.05) higher clay content and lower Db than the soils at hilltops
for C sequestration in croplands in this region. Irrespective of being at
and middle slopes. The soils were classified as Typic Udorthents and
different topographic positions, equivalent C stocks in hilltop and
Udic Haplustepts (USDA, 1999).
grasslands soils probably represent steady state values for soils in the
experimental area.
3.2. Total organic carbon Soils at the eroding slopes experienced the greatest loss of organic C,
amounting to ~58% of TOC stored in non-eroded hilltop soils. The loss
Soils at hilltops and the backslope grasslands had equivalent TOC of organic C from eroding slopes may be attributed to detachment of
concentrations, which were significantly (p < 0.05) higher than the particles and breaking of soil aggregates leading to exposure of pro-
cropland and the middle soils (Table 2). Soils at the middle slopes had tected organic C inside the aggregates. This resulted in enhanced rate of
the lowest concentration of TOC (1.78 ± 0.13 g kg−1), which was decomposition and loss of unprotected organic C during the detachment
lower by 53 to 61%, compared with the soils at other topographic and transport processes. This contention is supported by our results on
positions. Similarly, TOC stocks were the highest in grassland and aggregate size distribution and aggregate associated C (see Section 3.5).
hilltop soils followed by croplands and the lowest in eroded middle Soils at the eroding slopes had the lowest proportion of macro-ag-
gregates and the minimum C density in both macro- and micro-ag-
Table 2 gregates. Though the proportion of micro-aggregates in eroding soils
Total organic carbon (TOC) concentration, stocks and labile C pools in surface
did not decrease compared to soils at other slope positions yet the C
soil (0–0.15 m) at hilltops, middle slope eroded soils, backslope grasslands and
density of micro-aggregates in eroded soils was much smaller. This was
croplands in Siwalik foothill region of northern India. Values in parenthesis
indicate standard error from mean. probably because macro- and micro-aggregates in non-eroded upslope
soils were broken apart during the detachment and transport process
Property Hilltop Eroded Grassland Cropland (erosion) resulting in the replacement of the pre-existing or old micro-
TOC (g kg )−1 c
4.32 (0.33) a
1.78 (0.13) c
4.56 (0.29) 3.75b (0.29) aggregates at middle slope positions by the recently broken macro-ag-
TOC stock (Mg ha−1) 10.1c(0.64) 4.2a (0.26) 10.1c (0.57) 8.4b (0.65) gregates, which had lost considerable amount of C because of enhanced
WEOC (mg kg−1) 27.6b (3.0) 12.1a (1.9) 28.8b (2.8) 23.8b (2.9) decomposition. Van Oost et al. (2007) suggested that replacement of
HWC (mg kg−1) 174b (15.3) 84a (10.7) 219c (12.9) 215c (13.3) TOC at sites of erosion and an increased C decomposition during the
CVL (g kg−1) 0.13a (0.02) 0.16ab (0.05) 0.38c (0.05) 0.25b (0.03)
CL (g kg−1) 0.40b (0.04) 0.21a (0.02) 0.61c (0.06) 0.47b (0.04)
detachment and transport process are the mechanisms that control the
CLL (g kg−1) 1.12a (0.08) 1.25b (0.09) 1.67c (0.10) 1.40b (0.10) source and/or sink behavior of the erosion process. Assuming that the
CR (g kg−1) 2.67c (0.23) 0.16a (0.01) 1.91b (0.13) 1.63b (0.14) accelerated erosion at the experimental sites has been taking place
KMnO4-C (g kg−1) 0.61b (0.05) 0.43a (0.02) 0.82c (0.03) 0.54b (0.03) during the last 50 years, an estimated 0.12 Mg C ha−1 year−1 is being
Non-labile C (g kg−1) 3.71c (0.26) 1.35a (0.09) 3.74c (0.29) 3.21b (0.21)
lost through soil erosion in the experimental region. Our estimate is
CMI 70.2c (5.23) 55.8a (3.92) – 62.4b (4.31)
within the range reported for SOC erosion rates of
Mean values for a soil property for different land-uses and eroded sites followed 0.02–0.218 Mg C ha−1 year−1 in different eco-systems (Albergel et al.,
by different letters are significantly different (p < 0.05) by Duncan's multiple 2006; Smith et al., 2005). Considerably high rate of C loss from middle
range test (DMRT). slopes emphasizes the need to control soil erosion. The revegetation of

175
P. Singh, D.K. Benbi
degraded slopes is one of the major strategies for the control of soil
erosion and recovery of these fragile ecosystems, which have been
disturbed by anthropogenic activities. Revegetation with grass in the
Loess Plateau of China have been reported to provide an effective
protection against soil erosion and reduce losses of soil organic C
through runoff and transport of sediments (Jia et al., 2012; Tang,
2004).
3.3. Labile pools of soil organic carbon
Water extractable organic carbon (WEOC) comprised a very small
proportion (0.63–0.68%) of TOC, and did not differ significantly among
hilltop, grassland and cropland soils. It was lower by 49–58% in soils at
middle slopes than the other three slope positions (Table 2). Hot water
soluble C comprised between 4.0 and 5.7% of TOC in different soils. Its
concentration in soils at middle slope was lower by 51.7–61.7% com-
pared to hilltop, grassland and cropland soils. Hot water soluble C
concentration did not differ significantly between grassland and crop-
land soils, which were significantly (p < 0.05) superior to hilltop soils.
Our results showing that land-use, grassland versus cropland both of
which existed at the lower end of the slope, did not influence WEOC
and HWC concentrations significantly, are in contrast with Chantigny
(2003), who reported higher concentration of water soluble C in soils
under grasslands compared to cropland soils. This was probably be-
cause of regular application of organic manure in croplands of our
study, which supplied much soluble organic C and deposition of water
soluble C from the eroding slopes (Benbi et al., 2015). This is supported
by our observation that the lowest concentrations of WEOC and HWC
occurred in soils at eroding slopes suggesting that most of the water
soluble C has been lost by erosion and because of lack of vegetation
there is no replenishment of water soluble forms of C through root
exudates. Higher concentration of HWC in soils at hilltops, than the
soils at other slope positions suggests that the organic matter is more
stable in soils at higher elevation than at the lower elevations probably
because of differences in vegetation and the quality of C returned to the
soil. The variation in HWC has been reported to be influenced by root
exudation pattern of crop species and quality of C input to the soil (Xu
and Juma, 1993).
Characterization of organic matter composition using a gradient of
oxidizing conditions showed that the grassland soils had the highest
concentration of very labile (CVL) and labile C (CL) among all the soils.
While hilltop, cropland and middle slope eroded soils did not differ
significantly from each other in CVL, but the eroded soils had sig-
nificantly lower concentration of CL than the other soils. Grassland soils
had the highest concentration of less labile C (CLL) followed by cropland
and eroded soils and those at the hilltops had the lowest concentration.
Hilltop soils had the highest concentration of CR, followed by grassland
and cropland soils and the least in middle slope soils (Table 2). Very
labile C and CL constituted a greater proportion of TOC in grassland
soils than all other soils (Fig. 2). In other soils, the abundance of CVL and
CL was in the order eroded > cropland > hilltop soils and crop-
land > eroded > hilltop soils, respectively. Hilltop soils embodied
greatest proportion of TOC in recalcitrant form, and the soils at the
eroding slopes had the lowest proportion, while grassland and cropland
soils were in-between. About 62% of TOC was stabilized as recalcitrant
C in the hilltop soils as opposed to 41.9 to 43.5% in grassland and
cropland soils. Soils at middle slopes had only about 9% of TOC oc-
curring as CR, a major proportion of TOC was recovered as less labile C.
The recalcitrance index of C (CR as percent of TOC) for grasslands was
41.9%, which was within the range reported for different grasslands
after acid hydrolysis of soil (Plante et al., 2006). Cheng et al. (2007)
reported that recalcitrance index of C in croplands under sorghum
cultivation varied between 47 and 56%, which was slightly higher than
that observed for maize-wheat system (43.5%) in the present study.
Middle slope soils had a very low recalcitrance index (9%), which
suggests that organic C in these soils is less stable and could be easily
176
Catena 166 (2018) 171–180
V. labile Labile Less labile Recalcitrant
100
c a b b
80
Carbon fraction (% of TOC)
60
40
a c b
b
20
b d c
a
c d b
0 a
Hilltop Middle slope Grassland Cropland
Fig. 2. Distribution of soil organic C fractions of varying lability (as % of total
organic C) in soils at different slope positions and land-uses in Siwalik foothills
region of northern India. Bars followed by the same letter for a given fraction
are not significantly different (p < 0.05) by Duncan's multiple range test.
lost if remedial measures are not taken. Higher recalcitrance of C at
non-eroding slope positions may be attributed to physical protection of
C associated with micro-aggregates. Doetterl et al. (2012) also observed
significantly higher mean residence time for C deposited down-slope,
relative to eroding positions.
KMnO4-oxidizable C comprised 14.1 to 24.2% of TOC, and its
concentration was highest in grassland soils and the lowest in middle
slope soils with hilltop and cropland soils, which were at par, in-be-
tween (Table 2). Non-labile C constituted a major proportion of TOC
(76–86%) across soils though its concentration was lowest in middle
slope eroded soils followed by croplands. The grassland and the hilltop
soils had the highest concentration of non-labile C. Computation of CMI
in reference to grassland soils showed that hilltop soils had the highest
CMI followed by croplands and the eroded soils. KMnO4-oxidizable C
has been suggested as a measure of labile C pool representing the or-
ganic constituents of easy oxidizability such as labile humic material
and polysaccharides and is reported to account for 5–30% of TOC (Blair
et al., 1995; Conteh et al., 1999). Similar to other published studies
(Blair et al., 1995, 2006), we also observed significantly higher con-
centration of KMnO4-C in soils under grasslands compared to croplands.
Lowest concentrations of KMnO4-C and non-labile C in eroded soils in
our study, suggest the loss of both forms of C as a result of soil erosion.
This method has been used to develop CMI as an indicator of soil C
rehabilitation. Higher CMI values indicate soil C rehabilitation and
smaller values suggest soils being degraded (Blair et al., 1995). In the
present study, the lowest CMI of eroded soils reiterates soil degradation
along non-vegetated slopes. Relatively higher CMI for croplands com-
pared to eroded soils indicate some soil rehabilitation but a value of
62.4% compared to grassland soils (=100) indicates scope for C se-
questration in these soils. Though the index has been proposed as
sensitive indicator of direction of C change in soil (aggrading or de-
grading), it provided unrealistically low CMI for hilltop soils. While
both grassland and hilltop soils had statistically similar concentrations
of total and non-labile C yet the method yielded only a C habilitation
index of about 70% for soils at hilltops. This indicates that the index
cannot be used as a measure of C rehabilitation. These results are si-
milar to those reported by Benbi et al. (2015) for rice-wheat system.
P. Singh, D.K. Benbi Catena 166 (2018) 171–180

Table 3
Cumulative C mineralization (Cmin), microbial biomass C (MBC), basal soil respiration (BSR), mineralization (qM), respiratory (qCO2), and microbial (qmic)
quotients of surface (0–0.15 m) soils from hilltops, middle slope eroded soils, backslope grasslands and croplands in Siwalik foothill region of northern India. Values in
parenthesis indicate standard error from mean.
Land-use Cmin (mg kg−1) MBC (mg kg−1) BSR (μg g−1 soil h−1) qM (μg CO2-C μg TOC−1) qCO2 (μg C–CO2 h−1 × μg biomass C−1) × 103 qmic

b b bc b a
Hilltop 261.8 (14.7) 122.3 (9.2) 0.161 (0.014) 0.062 (0.002) 1.36 (0.136) 0.028a (0.0005)
Eroded 211.1a (11.7) 51.6a (6.0) 0.101a (0.013) 0.119d (0.004) 1.96b (0.247) 0.029a (0.0010)
Grassland 241.9ab (13.5) 125.1b (8.1) 0.143b (0.014) 0.054a (0.001) 1.16a (0.113) 0.027a (0.0004)
Cropland 278.5c (15.7) 104.8b (8.9) 0.180c (0.015) 0.076c (0.002) 1.78b (0.156) 0.028a (0.0010)

Mean values within a column followed by different letters are significantly (p < 0.05) different by Duncan's multiple range test (DMRT).

3.4. Mineralizable carbon, microbial biomass and microbial indices
After 45 days of incubation cumulative amount of C mineralized
was significantly higher in cropland soils at the bottom of the slope
compared with soils at other slope positions (Table 3). Cumulative
amount of CO2-C evolved from the grasslands and the hilltop soils did
not differ significantly. Microbial biomass C which comprised between
2.74 and 2.90% of TOC was significantly lower in soils at eroding slopes
than at other slope positions, which did not differ significantly among
themselves. Higher MBC in soils from non-eroding slope positions
(hilltop, grassland and croplands) may be attributed to the substrate C
availability through root biomass and root exudates in these soils
compared to un-vegetated soils at eroding slopes. Eroded soils exhibited
significantly lower basal soil respiration (BSR) than the cropland soils.
Conversely, the mineralization quotient (qM) was the highest in soils
from eroded slopes and the lowest in grasslands, whereas hilltop and
cropland soils had intermediate levels. The metabolic or respiratory
quotient (qCO2), which is an indicator of age structure of the microbial
biomass was significantly higher for eroded soils compared to grassland
and hilltop soils, but was at par with cropland soils. A qCO2 value close
to 2.0 for eroding soils and croplands indicate energetically less effi-
cient microbial community in these soils (Anderson, 2003). Higher
Cmin and BSR in croplands soils could be due to the application of
organic manure, which influences microbial population and activity
through changes in substrate supply.
3.5. Aggregate size distribution and aggregate associated carbon
Total water stable aggregates (WSA) did not differ significantly in
soils at non-eroding slope positions viz. hilltops, grasslands and crop-
lands, but were significantly (p < 0.05) lower at middle slopes
(Table 4). The WSA comprised 67.2–72.5% of soil, out of which
38.7–46.5% occurred as macro-aggregates and 25.1–28.4% as micro-
aggregates. Grassland and cropland soils had significantly (p < 0.05)
higher proportion of macro-aggregates and lower proportion of micro-
aggregates than the soils at hilltop and middle slopes. Relative
distribution of macro-aggregates within different size classes showed
the dominance of meso-aggregates (MesoA) (0.25 to < 2.0 mm) com-
pared to coarse macro-aggregates (CMacA, > 2.0 mm) in all the soils.
Among the MesoA, the 0.25–0.50 mm sized fraction was greatest in
amount, and those between 1.0 and 2.0 mm size were least in amount.
Among micro-aggregates, coarse micro-aggregates (CMicA) of size
0.11–0.25 mm were most abundant and fine micro-aggregates (FMicA)
of size 0.053–0.11 mm were least abundant (Table 4). Grassland soils
had the highest aggregate ratio and eroded soils had the lowest ratio.
Aggregate ratio did not differ significantly in hilltop and cropland soils.
It was higher by 8.5, 32.6 and 28.4% in the hilltop, grassland and
cropland soils, respectively than the soils at eroded sites. The MWD of
grassland soils was higher by 12.5% compared to eroded soils, but was
at par with hilltop and cropland soils. The MWD of hilltop and cropland
soils was higher by 9.1 and 17.8%, respectively compared to the eroded
soils.
Organic C concentration in CMacA was higher than the whole soil
irrespective of the slope position (Table 5). Among MesoA, organic C
concentration was generally higher in the 1.0–2.0 mm sized aggregates,
followed closely by 0.5–1.0 mm sized aggregates. The smaller-sized
MesoA (0.25–0.50 mm) had the lowest concentration of organic C.
Comparison of macro- and micro-aggregates showed that organic C
concentration was higher in macro- than micro-aggregates. The CMacA-
C concentration was significantly lower in soils at eroded slopes; and
the other three slope positions did not differ significantly. Similarly, the
organic C concentration among MesoA of different size fractions was
significantly lower in soils at eroded slopes than at other elevations.
Among all the soils, grasslands had the highest MesoA-C.
The CPC of WSA was significantly higher in soils under grasslands in
contrast to eroded slopes, which had the lowest CPC (Fig. 3). The
hilltop and cropland soils, however, did not differ significantly with
regard to CPC of WSA. Aggregates of different sizes also differed in CPC
(Fig. 4). In all the aggregate sizes, CPC was considerably lower for soils
at eroding slopes, compared with other slope positions. Regardless of
slope position, CMacA (> 2.0 mm) had the lowest and CMicA
(0.11–0.25 mm) the highest CPC. In general, CPC of aggregates

Table 4
Aggregate size (mm) distribution, aggregate ratio and mean weight diameter (MWD) in surface soil (0–0.15 m) at hilltops, middle slope eroded soils, backslope
grasslands and croplands in Siwalik foothill region of northern India.
Land-use Total water Macro-aggregates (%) Macro- Micro-aggregates (%) Micro- Aggregate ratio MWD (mm)
stable aggregates aggregates
aggregates CMacA¶ MesoA† (%) CMicA‡ FMicA♠ (%)
(%)
> 2.0 mm 1.0–2.0 mm 0.5–1.0 mm 0.25–0.50 mm 0.11–0.25 mm 0.053–0.11 mm

b ab c b ab b a
Hilltop 71.1 2.38 9.12 13.1 18.2 42.8 24.3 4.04b 28.3c 1.53ab 0.551ab
Eroded 67.2a 2.04a 6.49a 11.8a 18.4a 38.7a 23.6a 4.84ab 28.4c 1.41a 0.505a
Grassland 71.1b 2.87b 8.00b 14.9c 20.2ab 46.0c 21.2a 3.93a 25.1a 1.87b 0.568ab
Cropland 72.5b 2.86b 10.0c 15.1c 18.5b 46.5c 22.8a 3.28ab 26.1b 1.81ab 0.595b

Mean values within a column followed by different letters are significantly different (p < 0.05) by Duncan's multiple range test (DMRT).
¶
CMacA = coarse macro-aggregates.
†
MesoA = meso-aggregates.
‡
CMicA = coarse micro-aggregates.
♠
FMicA = fine micro-aggregates.

177
P. Singh, D.K. Benbi Catena 166 (2018) 171–180

Table 5
Aggregate associated C in surface soil (0–0.15 m) at hilltops, middle slope eroded soils, backslope grasslands and croplands in Siwalik foothill region of northern
India.
Land-use Aggregate associated carbon (g kg−1 aggregate)

CMacAC¶ MesoAC† CMicAC‡ FMicAC♠

> 2.0 mm 2.0–1.0 mm 1.0–0.5 mm 0.50–0.25 mm 0.25–0.11 mm 0.11–0.053 mm

c c b c b
Hilltop 5.10 4.87 4.76 4.58 4.08 3.86c
Eroded 2.10a 2.10a 1.95a 1.90a 1.71a 1.56a
Grassland 5.66c 5.20c 5.30b 4.84c 4.11b 3.95c
Cropland 4.65b 4.26b 4.39b 3.98b 3.55b 3.25b

Mean values within a column followed by different letters are significantly different (p < 0.05) by Duncan's multiple range test (DMRT).
¶
CMacAC = coarse macro-aggregate associated C.
†
MesoAC = meso-aggregate associated C.
‡
CMicAC = coarse micro-aggregate associated C.
♠
FMicAC = fine micro-aggregate associated C.

4.0 c and cropland soils was higher than the eroded slopes. In contrast, the
Carbon preservation capacity

b proportion of micro-aggregates in soils under grasslands was lower than

b the eroded soils. Higher proportion of macro-aggregates in grassland
3.0 soils compared to the eroded soils could be attributed to the addition of
root biomass C coupled with minimum disturbance due to absence of
(g kg-1 soil)

tillage under grassland soils. In the croplands, C input as plant biomass

(roots and leaf plus stubble) and manures could be the possible reason
2.0
for higher proportion of macro-aggregates, compared with the eroded
a soils. Application of organic manure causes formation of SOM asso-
ciations with silt plus clay particles and with micro-aggregates in the
1.0 first step, and then lead to the formation of macro-aggregate, when the
SOM binding capacity of the silt plus clay fractions is saturated
(Hassink, 1997; Tisdall and Oades, 1982). Organic matter stabilizes
0.0 aggregates by increasing the cohesion of aggregates through the
Grassland Cropland Hilltop Middle binding of mineral particles by organic polymers, and through the
slope physical enmeshment of particles by fine roots or fungi (Tisdall and
Oades, 1982).
Fig. 3. Carbon preservation capacity (CPC) of total water stable aggregates
Land-use change results in destruction and reformation of soil
(WSA) in the surface soil (0–0.15 m) of soils at different slope positions. Mean
values for different slope positions followed by same letter are not significantly structure that promotes C mineralization (Golchin and Asgari, 2008).
different (p < 0.05). Line bars represent standard error. The C storage in aggregates depends upon C input, rate of soil organic
matter decomposition and climatic conditions (Johnston et al., 2009).
Six et al. (2002) reported that organic C and stable aggregates mutually
1.2 Hilltop
protect each other as C is physically protected by its association with
Middle slope
Carbon preservation capacity

soil primary particles in aggregates. At the same time aggregate stabi-

1.0 Grassland
lity is enhanced by this association (Six et al., 2002). Among different
Cropland
sized aggregates, CMacA (> 2.0 mm) had higher TOC than any other
0.8
(g kg-1 soil)

aggregate class, showing the importance of macro-aggregates for C

sequestration (Dorodnikov et al., 2009). In this study, C concentration
0.6 in macro-aggregates (> 0.25 mm) was much higher than in the micro-
aggregates (< 0.25 mm). Among MesoA, C concentration decreased
0.4 with the size of aggregates. Ashagrie et al. (2007) also observed that
land-use and management changes affected MacAC more than MicAC.
0.2 Several researchers have reported that macro-aggregates under dif-
ferent land-use and soil management conditions were C-rich compared
0.0 to micro-aggregates (Benbi et al., 2016; Cambardella and Elliott, 1993;
0.11-0.25
>2.0

1.0-2.0

0.5-1.0

0.25-0.50

0.053-0.11

Puget et al., 1995).

3.6. Discriminant function analysis

Aggregate size (mm) classes
Fig. 4. Carbon preservation capacity (CPC) of different sized aggregates in the Two significant canonical discriminant functions involving MacA,
surface soil (0–0.15 m) at hilltop, middle slopes and under grassland and MWD, TOC and CR could group soils from different slope positions
cropland. Line bars represent standard error. though the soils from grasslands and croplands were not clearly dis-
tinguished (Fig. 5). But non-eroded soils at hilltops and those at the
increased as the macro-aggregates became finer (Fig. 4). The results eroding slopes were clearly separated. The cross-validated classification
showed that total WSA were lower at eroding slopes than the other showed that overall 98.8% of soils were accurately classified. Sig-
slope positions. However, the proportion of macro-aggregates in hilltop nificant canonical correlations were observed among the variable and
first two functions (0.964**, 0.749**, p < 0.01).

178
P. Singh, D.K. Benbi
Fig. 5. Plot of canonical discriminant functions for grouping soils at hilltop,
middle slopes, and under grassland and cropland in Siwalik foothills region of
northern India.
4. Conclusion
The study explored the interactive effects of topographic position
and land-use on quantity and composition of soil organic C along ty-
pical hill-slopes. Total and labile pools of soil organic C were sig-
nificantly influenced by topographic position and land-use. Soils at the
hilltop exhibited the highest stocks of total organic C, almost two-thirds
of which was stabilized in recalcitrant form. On the contrary, soils at
the middle slopes had the lowest stocks of TOC, which were pre-
dominantly (> 90%) in labile form suggesting that at middle slope
positions, soil organic C was amenable to further loss. Because of the
action of erosive processes and loss of organic C, the bare soils at middle
slopes had the lowest proportion of macro-aggregates and minimum C
density in both macro- and micro-aggregates. At the non-eroding
backslope positions, grassland soils had about 20% higher TOC stocks
compared to cropped bottomlands probably because of greater plant-
mediated C return to the soil and deposition of moved C from the
eroding upper slopes to grasslands. Prevalence of relatively high TOC
stocks in soils under grassland and at hilltops suggests that there is a
great potential for C sequestration at middle and backslope positions
through improved management and control of soil erosion.
Acknowledgement
This research was supported by the Indian Council of Agricultural
Research National Professor project.
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