Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci.

(Oct–Dec 2013) 83(4):513–524

DOI 10.1007/s40011-013-0162-1

RESEARCH ARTICLE

Urease Activity in Various Agro-ecological Sub-regions of Black

Soil Regions of India
Kulandaivelu Velmourougane • M. V. Venugopalan • T. Bhattacharyya •

Dipak Sarkar • D. K. Pal • Apeksha Sahu • P. Chandran • S. K. Ray •

Champa Mandal • K. M. Nair • Jagdish Prasad • R. S. Singh •
Pramod Tiwary

Received: 24 September 2012 / Revised: 4 January 2013 / Accepted: 22 January 2013 / Published online: 12 February 2013
Ó The National Academy of Sciences, India 2013

Abstract A study was undertaken in the established available water content. Electrical conductivity, calcium
benchmark soil series in different agro-ecological sub-regions carbonate and cation exchange capacity showed negative
of Black Soil Regions of India with the objective to assess the correlation in chemical properties at all the soil depths.
urease activity as a function of soil depth, bio-climate, crop-
ping system and land use type. The urease activity declined Keywords Urease activity
Bio-climates

with increase in soil depth. Maximum activity was restricted Cropping systems
Agricultural land use

within 0–30 cm of soil depth. Cropping systems and bio-cli- Principal Component Analysis
mates significantly (p \ 0.01) influenced the urease activity in
soil. The average urease activity in different bio-climates was
in decreasing order viz. sub-humid (moist) [ sub-humid Introduction
(dry) [ semi-arid (dry) [ arid. The activity in different
cropping systems was in decreasing order viz. legume- Soil enzymes have been suggested as one of the potential
[ sugarcane- [ cereals- [ cotton-based cropping system. biological indicators of soil quality because of their rela-
Higher urease activity was observed in irrigated agro-systems tionship to soil biology, ease of measurement, and rapid
as compared to the rainfed agricultural systems. High man- response to changes in soil management [1]. Among vari-
agement practices increased urease activity as compared to ous soil enzymes, urease (urea amidohydrolase EC [Link]:
low management. In physical properties, urease activity was catalyzing the hydrolysis of urea to CO2 and NH3) is very
negatively correlated with sand, fine clay, bulk density and widely distributed in nature, and has been detected in
plants, animals, and microorganisms [2, 3]. Urease plays an
important role in the efficient use of urea fertilizer in soil
and the changes in urease activity can be used as an indi-
K. Velmourougane (&)
M. V. Venugopalan
A. Sahu rect indicator of the variation in the pool of potentially
Central Institute for Cotton Research, Nagpur, available N in a soil [4]. Urease activity influences the
Maharashtra, India optimum use of urea fertilizer, N volatilization, N leaching
e-mail: velicar@[Link]
and environmental pollution related to N [5, 6]. While, low
T. Bhattacharyya
D. Sarkar
D. K. Pal
P. Chandran
urease activity might cause added urea to be lost by
S. K. Ray
C. Mandal
J. Prasad
P. Tiwary leaching; on the other hand, a higher activity might result
National Bureau of Soil Survey and Land Use Planning, in excessive hydrolysis of added urea and subsequently
Nagpur, Maharashtra, India
ammonia can be lost by volatilization [7]. Li et al. [8]
K. M. Nair reported that urease activity was closely related to soil
Regional Centre, National Bureau of Soil Survey and Land Use nutrient conditions and recommended that urease should be
Planning, Bangalore, Karnataka, India considered as an important parameter for estimating the
Soil Quality Index.
R. S. Singh
Regional Centre, National Bureau of Soil Survey and Land Use In India, though a few studies have been reported about
Planning, Udaipur, Rajasthan, India the soil urease activity [9–11], information on the urease

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514 K. Velmourougane et al.

Table 1 Characteristics of selected BM spots in BSRs of India [13]

AESR Bio-climates MAR (mm) Soil series MSL (m) Districts States Soil sub-group classification

6.1 Arid 520 Nimone 517 Ahmednagar Maharashtra Sodic Haplusterts

5.1 Arid 533 Sokdha 25 Rajkot Gujarat Calcic Haplusterts
8.1 SAd 612 Coimbatore 421 Coimbatore Tamil Nadu Vertic Haplustepts
3.0 SAd 632 Teligi 379 Bellary Karnataka Typic Haplusterts
6.4 SAd 638 Acchamatti 573 Dharwad Karnataka Sodic Haplusterts
7.1 SAd 650 Nandyal 212 Kurnool Andhra Pradesh Chromic Haplusterts
5.1 SAd 650 Bhola 76 Rajkot Gujarat Typic Haplusterts
8.3 SAd 660 Kovilpatti 81 Tuticorin Tamil Nadu Gypsic Haplusterts
8.2 SAd 661 Siddalaghatta 717 Kolar Karnataka Vertic Haplustepts
7.2 SAd 764 Kasireddipalli 538 Medak Andhra Pradesh Sodic Haplusterts
6.2 SAd 789 Vasmat 372 Hingoli Maharashtra Sodic Haplusterts
6.3 SAd 794 Paral 267 Akola Maharashtra Sodic Haplusterts
5.2 SHd 1053 Sarol 564 Indore Madhya Pradesh Typic Haplusterts
10.3 SHd 1100 Ghulguli 509 Shahdol Madhya Pradesh Typic Haplusterts
10.2 SHm 1127 Panjri 309 Nagpur Maharashtra Typic Haplusterts
10.1 SHm 1209 Nabibagh 501 Bhopal Madhya Pradesh Typic Haplusterts
7.3 SHm 1250 Tenali 15 East Godavari Andhra Pradesh Halic Haplusterts
AESR agro-ecological sub-regions, MAR mean annual rainfall (mm), Arid \550 mm, SAd semi-arid dry (850–550 mm), SHd sub-humid dry
(1,100–1,000 mm), SHm sub-humid moist ([1,100 mm), MSL elevation above mean sea level

activity in the Black Soil Regions (BSRs) of India which two representative pedons/soil profile (under the same soil
covers about 76.4 M ha of total geographical area of the series) were included—one under low management (LM)
country is lacking. Hence, a survey was undertaken in the which is characterized by application of low NPK, rarely
established benchmark (BM) soil series of BSR of India applied manures, removal of residues and biomass, no soil
with the objective to assess the urease activity as a function moisture conservation practices and the other under high
of soil depth, bio-climate, cropping system and land use management (HM) which is characterized by application of
type. This is the first extensive report on urease activities in higher NPK, regular application of organic manures,
BSR of India and the information generated through this incorporation of residues, adoption of soil moisture con-
study will be highly useful for the refinement and man- servation techniques (ridge furrows, bunding, broad bed
agement of nitrogen fertilization to crops and also for the and furrow).
assessment of land quality in BSR of India specifically.
Soil Physico-chemical and Microbiological Analysis

Material and Methods The international pipette method was applied for particle-
size analysis for quantifying the sand, silt and clay
Site Description and Sampling fractions according to the size segregation procedure of
Jackson [14]. Bulk density (BD) was determined by field-
The characteristics of selected BM spots of BSR of India moist method using core samples (diameter 50 mm) of
are summarized in Table 1 and the cropping systems and known volume (100 ml) [15, 16]. Hydraulic conductivity
management practices adopted in the BM spots are pre- was measured by taking 200 g of soil, uniformly tapped
sented in Table 2. The soil samples (approximately 1 kg and saturated overnight. It was measured by taking an
each from different horizons of a pedon) were collected hourly observation until three constant observations were
from the representative BM spots in the BSR of India obtained. It was measured in cm h-1 [17]. The chemical
(Fig. 1) covering specific bio-climatic systems in six AERs characteristics of soil were determined by standard proce-
(agro-ecological regions) and 17 AESRs (agro-ecological dures [18]. For microbiological analysis, soil samples
sub-regions—3.0, 5.1, 5.2, 6.1, 6.2, 6.3, 6.4, 7.1, 7.2, 7.3, collected at different soil depths from different BM spots
8.1, 8.2, 8.3, 10.1, 10.2, 10.3, and 5.1) [12] accounting for were serially diluted in 90 ml Ringers solution up to 10-4
19 % (117 M ha) [13] of total geographical area of the dilution and 1 ml of aliquot was pour plated in selective
country. The soil series were selected in such a way that in media (Nutrient Agar for bacteria [19], Martin’s Rose
any agricultural system under a particular cropping pattern, Bengal Agar for fungi [20], Ken Knights and Munaier’s

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Soil Urease Activity, Black Soil Regions of India 515

Table 2 Cropping systems and management practices adopted in selected BM spots in BSRs of India
BM spots HM LM
Cropping systemsa Agriculture Cropping systems Agriculture

Nimone Soybean–wheat/chick pea Irrigated Soybean/pearl millet/chick pea Irrigated

Sokdha Cotton ? green gram/pearl millet Rainfed Cotton ? green gram/pearl millet/sorghum Rainfed
Coimbatore Maize–chick pea Irrigated Single cropping of chick pea Rainfed
Teligi Triple cropping of rice Irrigated Maize/sorghum–chick pea Rainfed
Acchamatti Cotton–wheat/safflower/sorghum Irrigated Maize–chick pea Rainfed
Nandyal Rice–rice Irrigated Cotton/sunflower Rainfed
Bhola Cotton–wheat Irrigated Cotton–wheat Irrigated
Kovilpatti Single cropping of sorghum Rainfed Single cropping of cotton/sunflower/chick pea Rainfed
Siddalaghatta Fruits crops ? sunflower/sorghum Irrigated Rice–maize–tomato Irrigated
Kasireddipalli Soybean ? pigeon pea/maize–sunflower Rainfed Chick pea/sorghum Rainfed
Vasmat Sugarcane Irrigated Rice-fallow Irrigated
Paral Cotton ? soybean/green gram ? sorghum Irrigated Cotton ? black gram/chick pea ? sorghum Irrigated
Sarol Soybean–wheat Irrigated Soybean–chick pea Irrigated
Ghulguli Pigeon pea/mustard/green gram Rainfed Rice–wheat/chick pea Irrigated
Panjri Single crop of cotton/soybean Rainfed Soybean–wheat/soybean–chick pea Rainfed
Nabibagh Soybean–wheat/soybean–chick pea Irrigated Soybean–wheat/soybean–chick pea Irrigated
Tenali Rice–rice Irrigated Rice–rice Irrigated
HM practices LM practices

Application of higher NPK Application of low NPK

Regular application of organic manures Manures rarely applied
Incorporation of residues Removal of residues and biomass
Adoption of soil moisture conservation techniques (ridge furrows, bunding, broad bed and furrow) No soil moisture conservation practices
a
Cropping systems: ‘/’ = or; ‘?’ = intercropping; ‘–’ = followed by

Agar [19] for actinomycetes and Buffered Yeast Agar for (Ag2SO4) and steam distillation of an aliquot of the resulting
yeast). The plates were incubated at optimum temperature soil suspension with MgO for 4 min.
(28 ± 1 °C for bacteria and yeast; 30 ± 1 °C for fungi and
actinomycetes) in triplicates. The microbial colonies Statistical Analysis
appearing after the stipulated time period of incubation
(3 days for bacteria and yeast; 5 days for fungi; 7 days for The data pertaining to BSR coming under different bio-
actinomycetes) were counted and expressed as total cul- climates, soil depths, cropping systems, land use, soil sub-
turable colony forming units (Cfus)/g of the sample. groups [22], and management practices were pooled
together and analysed for descriptive statistics, ANOVA,
Assay of Urease Activity in Soil and principal component analysis using statistical soft-
ware’s SAS version 9.2 and JMP-8.
Urease (EC [Link]) activity in selected BM soils (air-dried,
crushed and sieved through a 2-mm mesh screen) was assayed
in duplicate by the method described by Tabatabai and Results and Discussion
Bremner [21], which involves the determination of the
ammonia released by urease activity when 5 g of soil was Variations in Urease Activity in BM Spots of BSR
incubated with 9 ml of 0.05 M Tris (hydroxymethyl) ami- of India: Soil Depth Influence
nomethane buffer (pH 9.0), 1 ml of 0.2 M of urea solution and
toluene at 37 °C for 2 h. The ammonia released was deter- The urease activity was found to decline in all the BM
mined by a procedure involving treatment of the incubated spots studied with increase in soil depth (Table 3). The
soil sample with 2.5 M KC1 containing a urease inhibitor maximum urease activity was recorded in the surface

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516 K. Velmourougane et al.

Fig. 1 Agro-ecological sub-regions of India and the locations of BM spots in BSRs of India

horizon (0–15 cm) and almost 50 % of urease activity was surface soil as compared to the deeper soil depths was the
found to be restricted within 0–30 cm in all the BM spots. result of greater availability of urea, organic C, nutrients
Higher urease activity was recorded in HM BM spots as and stimulated microbial activity in the surface soil [23–
compared to LM spots and the urease activity differed 25]. Depth of root penetration and nitrogen exhaustive
significantly (p \ 0.01) between the BM spots. Among the characteristics of crops may also be another reason for the
BM spots in HM, highest urease activity was recorded in decline of urease activities in deeper soil layers. The low
Nimone soil series of Maharashtra (47.5 lg NH4?-N g-1), content of total nitrogen in soil due to cultivation of high-
while the lowest urease activity was recorded in Bhola exhaustive crops together with low soil organic carbon
series of Gujarat (2.6 lg NH4?-N g-1) at 15 cm soil depth. content was also reported as one of the reasons for lesser
In LM, highest urease activity was recorded in Vasmat soil urease activity in sub-surface [26]. Urease activity was
series of Maharashtra (44.7 lg NH4?-N g-1), while the reported to be proportional to organic C distribution in each
lowest urease activity was recorded in Nandyal series of soil profile. Its maximum activity was concentrated in the
Andhra Pradesh (2.0 lg NH4?-N g-1) at 15 cm soil depth. surface soil which decreased with depth [23, 24]. The sharp
The major reason for increased urease activity in the decline in urease activity with increasing soil depth have

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Soil Urease Activity, Black Soil Regions of India 517

Table 3 Urease activity (lg NH4?-N g-1 dry soil) in selected BM spots of BSR of India
BM soil series Soil depth (cm)
0–15 15–30 30–50 50–100 100–150
HM LM HM LM HM LM HM LM HM LM

Nimone 47.5 42.4 48.0 30.4 39.6 23.4 26.6 18.1 20.8 13.6
Sokdha 8.00 5.40 7.10 5.10 5.60 4.70 4.40 4.00 – –
Coimbatore 38.4 25.8 30.0 23.1 26.2 20.9 18.9 15.2 16.0 11.4
Telgi 13.0 9.60 8.50 8.30 6.30 6.80 4.20 5.20 3.00 4.00
Acchamatti 14.6 2.30 12.7 7.70 11.4 11.3 7.80 11.3 5.70 9.10
Nandyal 3.40 2.00 3.20 2.40 2.90 2.50 2.20 2.10 1.70 1.60
Bhola 2.60 ND 2.20 ND 2.10 ND 1.80 ND – ND
Kovilpatti 10.7 5.90 9.90 4.80 8.80 3.70 6.20 2.30 4.50 1.70
Siddalghatta 16.4 17.6 16.6 15.5 15.7 12.1 11.4 7.40 8.90 5.40
Kasireddipalli 25.6 21.8 11.2 16.7 9.30 14.0 6.50 10.4 – –
Vasmat 13.6 44.7 16.4 32.9 14.1 26.8 9.60 16.8 7.60 12.1
Paral 20.6 10.0 12.9 6.10 11.0 5.20 8.60 4.20 6.30 3.90
Sarol 38.9 34.2 20.2 24.7 14.3 17.9 8.50 12.5 6.40 10.1
Ghulghuli 8.60 8.30 7.50 9.00 6.90 8.20 6.60 – 5.90 –
Panjari 16.1 15.9 15.5 14.9 13.1 12.3 9.10 8.10 6.50 5.80
Nabibagh 42.5 35.3 39.7 29.0 29.6 24.5 19.3 18.8 15.0 15.3
Tenali 3.60 2.20 3.40 1.90 2.90 1.80 2.50 1.80 2.20 1.50
Standard deviation 14.4 14.6 12.7 10.5 10.2 8.27 6.73 6.01 5.58 4.83
Coefficient of variation 75.6 82.9 81.7 72.3 78.9 67.5 74.2 65.3 70.7 65.8
Standard error of mean 3.49 3.67 3.09 2.62 2.47 2.06 1.63 1.55 1.49 1.34
p value 0.003** 0.001*** 0.002** 0.005** 0.001** 0.006** 0.005** 0.003** 0.002** 0.009**
HM high management, LM low management, ND not determined
**, *** p values significant at 0.01 and 0.001 probability level respectively

Table 4 Bio-climates and soil sub-groups on urease activity (lg NH4?-N g-1 dry soil) in BSRs of India
Soil depth (cm)
0–15 15–30 30–50 50–100 100–150

Bio-climates
SHm 19.3b 17.4a 14.0a 9.9a 7.7a
a b b c
SHd 22.5 15.3 11.8 6.9 5.6c
SAd 15.7c 12.7c 11.1b 8.0b 6.4b
d d c d
Arid 6.7 6.09 5.14 4.2 2.9d
Soil sub-groups
Vertic Haplustepts 24.6a 21.3a 18.7a 13.2a 10.5a
a b b b
Sodic Haplusterts 24.3 19.5 16.6 12.0 7.90b
Typic Haplusterts 18.8b 14.9c 11.8d 7.80d 6.00c
Gypsic Haplusterts 16.6c 14.6c 12.5c 8.50c 6.20c
d d e c
Calcic Haplusterts 13.4 12.2 10.3 8.40 –
Halic Haplusterts 5.80e 5.30e 4.70g 4.30e 3.70d
f e f e
Chromic Haplusterts 5.40 5.50 5.40 4.30 3.30d
Bio-climate and soil sub-groups with the same letter in the columns are not significantly different following Tukey HSD (0.01)

been reported due to either the lower microbial activity cultivation of legumes are also reported to be the important
[25], and limited availability of organic substrates [27]. factors which increase the mineral nitrogen in soil resulting
Application of higher nitrogenous fertilizers and in increased urease activity in the surface horizon [26].

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518 K. Velmourougane et al.

Table 5 Cropping systems on urease activity (lg NH4?-N g-1 dry soil) in BSRs of India
Cropping systems Crops Soil depth (cm)
0–15 15–30 30–50 50–100 100–150

Legume-based system Chick pea 47.4a 40.1a 35.1a 26.0a 11.6b

c c c c
Soybean 35.4 27.4 21.5 14.9 12.6a
Pigeon pea 8.8f 7.5f 7.2f 6.7f 0.0
b b b b
Sugarcane-based system Sugarcane 44.3 33.1 26.9 17.0 12.3a
d d d d
Cereal-based system Maize 22.3 19.2 17.7 13.7 11.3b
e e e e
Sorghum 13.7 13.5 12.5 9.0 6.8c
f f fg g
Rice 8.7 7.8 6.7 4.7 3.7e
Cotton-based system Cotton 9.3f 7.6f 6.6g 5.0g 4.5d
Cropping systems with the same letter in the columns are not significantly different following Tukey HSD (0.01)

Table 6 Land use types and management levels on urease activity (lg NH4?-N g-1 dry soil)
Soil depth (cm)
0–15 15–30 30–50 50–100 100–150

Agricultural land use

Irrigated 24.0a 19.0a 15.3a 10.8a 8.7a
b b b b
Rainfed 12.5 10.3 9.1 7.0 5.6b
Management level
HM 19.1a 15.7a 12.9a 9.0ns 7.9ns
LM 17.6b 14.5b 12.3b 9.2ns 7.3ns
ns non significant
Land use types and management levels with the same letter in the columns are not significantly different following Tukey HSD (0.01)

Bio-climates and Soil Sub-groups on Urease Activity It is well known that the hydrolyzing activity of urease
follows the Michaelis–Menten kinetics, and it is influenced
The pooled analyses of urease activity in different bio-climates by several factors including the soil type, organic matter
and soil types of BM spots (inclusive of cropping systems, content, temperature, soil moisture, pH and the amount of
management, and land use effects) indicated significant dif- added nitrogen [28–31]. Urease activity was related to
ferences (p \ 0.01) between the bio-climate, soil type as well seasonal changes in soil temperature and moisture [32] and
as the soil depth (Table 4). At the surface layer (0–15 cm) reported to show considerable sensitivity to water avail-
higher urease activity was recorded in sub-humid (dry) (SHd) ability [33], however, the activity of urease has not always
(22.5 lg NH4?-N g-1) with a range between 8.6 and 38.9 lg been reported as correlated with soil water availability
NH4?-N g-1 followed by sub-humid (moist) (SHm) (19.3 lg [34]. Urease activity is also reported to increase as the
NH4?-N g-1, range = 3.6–42.5 lg NH4?-N g-1) and the temperature rises [35]. The variation in urease distribution
least urease activity was recorded in arid regions (6.7 lg NH4?- in different soil types is reported to be influenced by
N g-1). The average urease activity in different bio-climates physical and chemical properties of the soil [9, 36, 37].
declined with the depth in decreasing order of SHm [
SHd [ semi-arid (dry) (SAd) [ arid. In soil sub-groups, Ver- Cropping Systems on Urease Activity
tic Haplustepts recorded higher urease activity (24.6 lg NH4?-
N g-1) followed by Sodic Haplusterts (24.3 lg NH4?-N g-1), Among the cropping systems effects at the surface layer
while the lowest urease activity was observed in Halic Hap- (0–15 cm) (inclusive of bio-climates, soil types, manage-
lusterts (5.8 lg NH4?-N g-1) and Chromic Haplusterts ment, and type of agriculture effects), legume-based
(5.4 lg NH4?-N g-1) at the surface soil (0–15 cm). The cropping system (chick pea/soybean/pigeon pea) recorded
average urease activity in different soil sub-groups were in higher urease activity (Table 5). In legume-based system,
decreasing order of Vertic Haplustepts [ Sodic Haplust- chick pea (47.4 lg NH4?-N g-1) followed by soybean
erts [ Typic Haplusterts [ Gypsic Haplusterts [ Calcic (35.3 lg NH4?-N g-1) recorded higher urease activity and
Haplusterts [ Halic Haplusterts [ Chromic Haplusterts. sugarcane-based cropping system 44.2 lg NH4?-N g-1)

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Soil Urease Activity, Black Soil Regions of India 519

Table 7 Correlation matrix for urease activity and soil parameters

Variables 0–15 cm 15–30 cm 30–50 cm 50–100 cm 100–150 cm

Sand (%) -0.308 -0.233 -0.235 -0.222 -0.068

Silt (%) 0.343 0.363 0.158 0.247 0.189
Clay (%) 0.083 0.013 0.155 0.059 -0.067
Fine clay (%) -0.163 -0.050 0.011 -0.102 -0.076
BD (Mg m-3) -0.229 -0.243 -0.195 20.405 20.395
1/3 bar 0.017 -0.128 -0.049 -0.117 20.409
15 bar 0.088 0.090 -0.153 -0.150 -0.308
1 bar -0.111 -0.259 -0.091 -0.086 -0.334
3 bar -0.163 -0.282 -0.128 -0.055 -0.292
-1
HC (cm h ) 0.455 0.204 0.131 -0.091 0.461
Water pH (1:2) 0.115 0.001 -0.142 -0.230 -0.158
KCl pH (1:2) 0.213 0.170 0.202 0.119 -0.024
EC (1:2) (dS m-1) -0.267 -0.031 0.241 0.228 -0.133
Organic carbon (%) 0.003 0.125 -0.100 -0.219 -0.123
Calcium carbonate (%) -0.019 -0.084 -0.057 -0.043 0.072
CEC -0.164 -0.250 -0.198 -0.223 -0.123
Base saturation (%) 0.148 0.007 -0.013 -0.049 -0.151
Exchangeable sodium percentage 0.325 0.381 0.267 0.188 0.068
Exchangeable magnesium percentage -0.163 0.097 0.182 0.257 0.162
Nitrogen (kg/ha) -0.072 0.085 0.050 0.243 0.109
Phosphorus (kg/ha) -0.132 0.166 -0.190 -0.203 0.049
Potassium (kg/ha) 0.159 -0.039 -0.043 0.028 -0.241
Microbial population (total Cfu) 0.154 0.030 0.064 0.087 0.019
In bold, significant values at the level of significance a = 0.050 (two-tailed test)

followed the legume system. The lowest urease activity significantly higher soil enzymatic activity in rotation of
was recorded in cotton-based cropping system (9.2 lg grains and legumes than in wheat fallow system. Urease
NH4?-N g-1). The urease activity showed significant dif- was also affected by plant types or plant species combi-
ference (p \ 0.01) between the cropping systems and the nations [41].
soil depths in all the BM spots. The average urease activity
in different cropping systems was in decreasing order of Agricultural Land Use and Management Practices
legume- [ sugarcane- [ cereal- and cotton-based crop- on Urease Activity
ping systems.
The higher urease activity in legume-based system The pooled analysis of soil urease data indicated significant
showed the contribution of legumes towards the greater differences (p \ 0.01) between the land use and the soil
availability of organic C and N and stimulated microbial depth. The data clearly indicates the dominance of irrigated
activity [38, 39]. Crop growth characteristics such as root agro-systems over the rainfed agricultural systems on the
growth, nitrogen fixation and utilization pattern were also urease activity (Table 6). The average urease activity in
governing higher urease activity in legume-based system. irrigated system was found to be 24 lg NH4?-N g-1, while
Availability of higher MN by way of huge nitrogenous in rainfed systems it was recorded as 12.5 lg NH4?-N g-1
fertilizer application and HM in sugarcane-based systems at the surface soil (0–15 cm). The pooled data on urease
reflected the impact of N fertilization on urease activity. distribution indicated significant differences (p \ 0.01)
The lesser urease activity in cereal- and cotton-based between the management practices and the soil depth. HM
cropping systems is mainly because of the crop charac- recorded higher urease activity (19.1 lg NH4?-N g-1) as
teristics (deep rooted and nutrient exhaustiveness) and compared to the LM (17.6 lg NH4?-N g-1).
management level (mostly rainfed with low inputs). The The average urease activity in irrigated system was
alteration of soil enzymatic activities resulting from culti- found to be almost 50 % higher than that of rainfed sys-
vation or differences in cropping system has been dem- tems. It is also clear that, the lesser urease activity under
onstrated by several researchers [40]. Khan [38] noted rainfed system is also contributed by the crops grown under

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Table 8 Contributions of the variables (%) after Varimax rotation

Variable 0–15 cm 15–30 cm 30–50 cm
F1 F2 F3 F4 F1 F2 F3 F4 F1 F2

Sand 10.33 2.71 5.43 0.05 9.35 5.05 1.96 0.00 11.06 0.30
Silt 0.29 14.12 10.71 0.69 0.56 14.98 9.13 0.18 0.07 1.27
Clay 13.02 1.66 0.00 0.22 12.85 0.04 0.42 0.10 12.68 0.03
Fine clay 4.75 0.00 10.29 0.16 6.04 1.45 0.13 2.02 6.18 0.09
BD 1.01 1.64 7.95 1.23 0.57 19.70 0.17 1.48 0.03 4.68
1/3 bar 12.02 2.87 0.01 0.98 12.60 0.03 0.18 0.26 13.19 0.71
15 bar 12.85 1.60 0.06 0.42 12.78 0.13 0.02 0.00 2.63 0.01
1 bar 12.83 1.13 1.18 0.81 13.16 0.30 0.13 0.82 14.37 0.00
3 bar 12.25 1.23 2.21 1.11 12.27 0.20 0.02 0.43 13.69 0.04
HC 0.07 0.66 16.71 0.44 0.56 0.96 0.54 27.00 1.69 0.21
Water pH 0.05 18.11 1.71 3.08 0.02 1.10 21.81 2.05 0.38 20.26
KCl pH 1.16 11.54 1.28 3.97 0.68 0.40 25.06 0.34 0.41 24.17
EC 1.16 0.06 14.03 14.73 0.04 3.51 8.60 1.51 0.22 1.92
OC 0.36 11.77 0.00 11.20 0.82 9.63 1.86 7.27 2.37 6.82
CaCO3 0.17 4.60 2.40 11.60 0.28 0.60 6.29 26.10 0.77 5.18
CEC 8.76 0.81 0.14 0.00 11.34 1.09 0.75 0.16 13.04 0.00
BS 3.53 5.67 3.99 0.26 2.24 1.98 3.51 0.85 3.63 0.22
ESP 0.17 0.34 0.85 13.28 0.01 9.40 10.15 4.13 0.06 13.83
EMP 0.00 0.00 0.01 2.93 0.11 0.24 2.69 0.36 0.25 0.48
N 0.03 15.73 0.22 7.04 0.06 10.70 1.25 2.59 0.39 14.51
P 0.06 1.77 0.26 22.07 0.22 0.00 1.09 21.20 0.05 0.94
K 1.59 0.02 1.54 1.28 1.94 0.44 0.09 0.95 1.77 3.19
Urease 0.02 0.00 18.98 0.00 0.15 16.73 0.35 0.06 0.02 1.08
Total Cfu 3.49 1.94 0.05 2.43 1.35 1.33 3.81 0.14 1.06 0.06
Percent variance (after Varimax rotation)
Eigenvalue 6.94 3.67 2.62 2.11 7.06 3.09 2.28 1.82 6.71 3.68
Variance (%) 27.75 15.26 11.32 9.59 28.80 10.25 11.73 8.57 26.14 13.18
Cumulative (%) 27.75 43.01 54.33 63.92 28.80 39.05 50.78 59.35 26.14 39.32
Variable 30–50 cm 50–100 cm 100–150 cm
F3 F4 F1 F2 F3 F4 F1 F2 F3 F4

Sand 0.02 3.50 9.39 1.16 0.14 0.46 10.02 1.19 0.04 1.99
Silt 1.67 22.14 0.72 3.83 0.02 1.65 0.02 1.71 0.49 6.20
Clay 1.13 1.32 12.63 0.04 0.08 0.03 10.94 0.03 0.09 0.12
Fine clay 6.01 4.51 8.91 0.30 1.45 1.02 7.46 0.46 0.83 0.00
BD 5.06 0.94 0.03 0.76 5.08 0.10 1.85 2.02 3.97 8.17
1/3 bar 0.46 0.01 12.34 0.95 0.00 1.55 10.06 1.25 1.66 4.25
15 bar 10.70 0.50 2.81 0.20 6.69 0.17 11.56 0.15 0.76 1.68
1 bar 0.49 0.49 13.04 1.54 0.01 1.95 10.38 1.60 3.86 1.87
3 bar 0.40 0.86 12.27 1.91 0.02 2.57 10.26 2.37 2.94 1.50
HC 12.22 0.55 4.51 1.16 9.31 3.33 2.05 3.47 5.07 15.13
Water pH 0.84 0.01 0.85 20.92 0.79 0.01 0.46 19.62 4.27 1.32
KCl pH 0.11 0.23 0.77 23.05 0.00 1.54 0.52 22.82 0.75 0.01
EC 0.12 15.68 0.11 0.83 0.32 27.28 0.03 0.93 30.09 0.04
OC 9.20 0.51 2.14 6.19 12.00 0.07 5.68 1.92 0.69 10.00
CaCO3 20.69 0.29 2.03 6.60 17.62 0.16 0.02 20.35 0.04 2.22
CEC 1.56 0.70 9.07 0.01 6.35 0.60 8.89 0.89 0.03 0.05

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Soil Urease Activity, Black Soil Regions of India 521

Table 8 continued
Variable 30–50 cm 50–100 cm 100–150 cm
F3 F4 F1 F2 F3 F4 F1 F2 F3 F4

BS 1.62 14.35 3.95 0.21 0.59 14.59 1.25 0.15 19.12 1.16
ESP 2.32 0.18 0.02 12.24 0.81 11.92 0.16 8.38 0.92 0.80
EMP 2.92 8.39 1.04 3.41 9.77 0.41 7.38 0.41 0.70 0.05
N 0.32 2.76 0.58 13.04 0.33 4.31 0.00 5.48 2.93 11.87
P 20.98 0.23 0.48 0.10 23.00 0.35 0.08 3.92 4.55 5.10
K 0.00 0.27 1.07 0.75 1.17 0.69 0.10 0.41 2.95 10.44
Urease 1.14 0.34 0.32 0.23 4.33 2.07 0.14 0.10 2.49 15.90
Total Cfu 0.02 21.24 0.92 0.57 0.12 23.18 0.69 0.36 10.78 0.12
Percent variance (after Varimax rotation)
Eigenvalue 2.78 2.03 6.87 3.26 2.84 2.11 7.50 3.24 2.50 1.92
Variance (%) 12.08 11.95 27.04 11.61 12.00 12.15 28.90 13.42 11.27 9.57
Cumulative (%) 51.39 63.35 27.04 38.65 50.65 62.80 28.90 42.33 53.59 63.17

such system. Rainfed system is dominated either by cereal- increased soil urease activity under irrigated cultivation
or cotton based which is managed with low inputs, while and concluded that urease activity was deeply influenced
the irrigated system (sugarcane- and legume based) is by water and heat factors. Impact of management practices
managed with high inputs. In BSR poor management on soil urease activity has been reported by several work-
coupled with low organic content makes the soil biologi- ers. Integrated application of vermicompost and nitrogen
cally inactive. Nourbakhsh and Monreal [42] reported fertilizers significantly increased soil urease activity [43].

a b c
(0-15 cm soil depth) (15-30 cm soil depth) (30-50 cm soil depth)

d e
(50-100 cm soil depth) (100-150 cm soil depth)

Fig. 2 Principle Component Analysis and factor rotation at different soil depths

123
522 K. Velmourougane et al.

Tillage practice influenced urease activity in the Ap hori- be obtained by measuring unculturable bacteria, as it is not
zon [44]. Nautiyal et al. [45] reported higher urease activity possible with the plate count technique. Similar results
in organically cultivated field which practiced no tillage were previously reported by Frankenberger and Dick [54]
and no removal of crop residues. Plant protection activities and by Reynolds et al. [55].
such as fungicide contamination of the soil are also
reported to significantly inhibit the urease activity [46].
Conclusion
Impact of Soil Properties on Urease Activity
Urease activity declined with increase in soil depth in all the
In the present study, though significant positive correlation
BM spots of BSR. The maximum urease activity was found to
was observed between urease activity and hydraulic con-
be in the surface horizon (0–15 cm) and almost 50 % of urease
ductivity at 0–15 and 100–150 cm, silt (15–30 cm) and
activity was restricted within 0–30 cm soil depth. Cropping
ESP (15–30 cm), urease activity was negatively correlated
systems and bio-climates significantly influence the urease
with sand, fine clay, bulk density (BD) and available water
activity in soil. The average urease activity in different bio-
content in soil, electrical conductivity (EC), calcium car-
climates was in decreasing order of SHm [ SHd [ SAd [
bonate and cation exchange capacity (CEC) at all the soil
arid, while its activity in different cropping systems was in
depths (Table 7). While physical parameters like clay and
decreasing order of legume [ sugarcane [ cereals [ cotton.
silt content showed positive correlation for urease activity
Higher urease activity was observed in irrigated agro-systems
in soil at all the depths, chemical parameters such as water
as compared to the rainfed agricultural system and HM
pH, organic carbon and base saturation showed positive
practice found to increase urease activity as compared to LM.
correlation only for the surface horizon (0–30 cm). In
On one hand India is spending huge capital on the import of
major nutrients, available nitrogen showed negative cor-
nitrogenous fertilizers while on the other hand enormous
relation for the surface and positive correlations for the
nitrogenous fertilizers are lost by N volatilization, N leaching
sub-surface horizons, while phosphorus and potassium
and environmental pollution. Hence better understanding of
does not follow any pattern. A weak positive correlation
the urease enzyme activity in soil of BSR will help in better
was observed between the total microbial population and
management of nitrogenous fertilizers to avoid soil pollution
urease activity at all the depths. At all the soil depths, soil
and increased cost of cultivation. This study will be highly
physical factors (water availability, sand, clay) were found
useful for the refinement and management of nitrogen fertil-
to influence the urease activity most followed by soil
ization to crops and also for the assessment of land quality in
chemical factors (pH, organic carbon and nitrogen)
BSR of India specifically.
(Table 8; Fig. 2a–e). CEC, base saturation and microbial
activity also influenced urease activity at all the depths. Acknowledgments This financial grant received from the World
The increase in soil water content favoured the micro- Bank sponsored ‘‘National Agricultural Innovation Project’’ (NAIP)
biological activity and subsequent increase in soil enzyme (Component-4: Indian Council of Agricultural Research) on
‘‘Georeferenced Soil Information System for Land Use Planning and
activities [47–49]. The difference in temperature and
Monitoring Soil and Land Quality for Agriculture’’ is gratefully
rainfall results in the variation of soil pH and EC [50], acknowledged. The authors are thankful to the Directors, CICR and
which show impacts on soil enzyme activity. In the present NBSS and LUP for extending facilities to carry out this research
study, soil pH was positively correlated with urease work.
activity. The close relationship between soil pH and
enzyme activity were reported by Deng and Tabatabai [51]
and Wang and Lu [52]. Rao and Ghai [2] reported non-
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