Environmental Pollution 134 (2005) 385–395

www.elsevier.com/locate/envpol

Amelioration of Indian urban air pollution phytotoxicity

in Beta vulgaris L. by modifying NPK nutrients
Anoop Singha, S.B. Agrawalb,*, Dheeraj Rathorea
a
Laboratory of Air Pollution and Global Climatic Change, Department of Botany,
Allahabad Agricultural Institute - Deemed University, Allahabad 211 007, India
b
Department of Botany, Banaras Hindu University, Varanasi 221 005, India
Received 28 November 2003; accepted 17 September 2004

Air pollution caused adverse impact on growth and biomass accumulation of Beta vulgaris L. plants
while higher fertility levels showed reduced yield losses.

Abstract

Air pollution levels are increasing at an alarming rate in many developing countries, including India and causing a potential
threat to crop production. Field experiments were conducted to examine the impact of urban air pollutants on biomass (yield) and
some physiological and biochemical parameters of palak (Beta vulgaris L. var. All Green) that grew from germination to maturity at
seven periurban sites of Allahabad city having different concentrations of air pollutants under different levels of nutrients. The 6 h
daily mean NO2, SO2 and O3 concentrations varied from 2.5 to 42.5, 10.6 to 65 and 3.5 to 30.8 mg mÿ3, respectively at different
locations. Levels of air pollution showed significant negative correlations with photosynthetic pigments, protein, ascorbic acid and
starch contents and catalase activity of palak leaves. A significant negative correlation was found for total biomass with SO2
(r Z ÿ0.92), NO2 (r Z ÿ0.85) and O3 (r Z ÿ0.91) concentrations. The increased fertilizer application (N, P and K) over the
recommended dose resulted in a positive response by reducing losses in photosynthetic pigments and total biomass. This study
proved that ambient air pollution of Allahabad city is influencing negatively to the growth and yield of palak plants.
Ó 2004 Elsevier Ltd. All rights reserved.

Keywords: Air pollution; Nutrients; Beta vulgaris; Biomass; Yield

1. Introduction concentrations ranged from 10.4 to 39.0 mg mÿ3 ppb in

The impacts of air pollution have long been recognized
as major cause of losses in crop production in several
developed countries. However, little attention has been
paid in developing countries, including India, on poten-
tial impacts of air pollution on growth and productivity.
The Indian national ambient quality data indicate that
emissions of a range of air pollutants are generally
increasing (Agrawal, 1998). The annual average of SO2
* Corresponding author. Tel.: C91 542 2368156; fax: C91 542
2368174.
E-mail address: sbagarwaldr@sancharnet.in (S.B. Agrawal).
most parts of the country, while NO2 concentrations were
found between 43.2 and 60.1 mg mÿ3 in metropolitan
cities. Pandey et al. (1992) reported elevated concen-
trations of O3 in Varanasi city, an adjoining district of
Allahabad, where significant negative influence of urban
air pollutants was recorded on a variety of plant species
growing in periurban areas (Agrawal et al., 2003).
Urban air pollution has direct impact on periurban
agriculture due to dispersion of pollutants in all
directions along the wind. During transportation
primary pollutants often form secondary pollutants,
causing greater adverse effects on crop production in
periurban areas. Effects of air pollutants have been

0269-7491/$ - see front matter Ó 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2004.09.017
386 A. Singh et al. / Environmental Pollution 134 (2005) 385–395
described in terms of foliar injury (Jacobson and Hill,
1970), reduction in photosynthetic pigments (Agrawal
et al., 1982), inhibition of physiological processes (Saxe,
1991), alteration in metabolic functions (Malhotra and
Khan, 1984), enzyme activities (Nandi et al., 1986) and
nutrient uptake and suppression of growth and yield of
agricultural crop plants (Lee, 2000; Verma et al., 2000;
Ribas and Penuelas, 2003; Singh et al., 2003).
Use of chemical protectants, such as growth regu-
lators, antioxidants and fertilizers is suggested to be
a short-term solution to reduce the risk of air pollution
damage. Researchers have suggested that application of
mineral nutrients promotes growth, and reduce pollut-
ant induced injury to crops (Ormrod et al., 1973).
Rajput and Agrawal (1994) have found that soybean
plants grown at recommended fertility levels were less
injured by SO2 in comparison to unfertilized crop.
In view of the above, the present investigation was
aimed to suggest an economical and ecofriendly solution
to ambient air pollution induced damage, by altering the
level of mineral nutrients in palak (Beta vulgaris var. All
Green) plants grown at different periurban and urban
sites of Allahabad city (Eastern Uttar Pradesh, India).
2. Materials and methods
The study was performed in the periurban and urban
environment of Allahabad city with a population of 0.85
million located in the eastern Gangetic plains of India
between 24  47#N latitude and 82  21#E longitude and
96 m above mean sea level. During the study period
mean minimum and mean maximum temperatures
ranged between 14.9–24.3  C and 30.8–34.6  C, respec-
tively (Table 1). The average relative humidity varied
between 59.8 and 68.4% and wind speed 3.5–6.8 km hÿ1.
Total precipitation was 164.2 mm during September and
77 mm during October (Table 1). Prominent wind was
westerly. The plant species chosen for this study is
a cheap and popular vegetable and consumed mainly as
a source of iron in the diet. Periurban area of Allahabad
provides 85% of the palak crop consumed in the city.
An experiment was conducted from September to
November 2001 at seven selected sites (viz. Allahabad
Agriculture Institute (AAI), Civil lines (CL), Mehdeori
(Mh), Jhunsi (Jh), Bahrana (Bh), Arail (Ar) and
Table 1
Meteorological data during the experimental period
Month and Precipitation Temperature (  C) Relative Wind
year (mm) humidity speed
Max. Min.
(%) (km hÿ1)
September, 164.2 34.6 24.3 68.4 6.8
2001
October, 2001 77.0 33.7 20.7 64.6 4.2
November, 2001 0.0 30.8 14.9 59.8 3.5
Rajrooppur (RRP)) on the periphery and within the
city of Allahabad. The location of sites and a brief
description of their characteristics are given in Fig. 1
and Table 2. The soil was prepared at one place by
mixing garden soil and farmyard manure in 3:1 ratio
following the normal agronomical practices for unifor-
mity of edaphic conditions. Soil used in the experiment
had pH 7.62, organic carbon 1.64%, N 690 mg 100 gÿ1
soil, P 16.4 mg 100 gÿ1 soil and K 136.2 mg 100 gÿ1.
Palak var. All Green seeds were sown in pots (30 cm
diameter) with four treatments of fertilizers, i.e. without
fertilizer (F0), recommended dose (RD) of N, P and K
(F1), one and half times of RD of N, P and K (F2) and
two times of RD of N, P and K (F3) on September 26,
2001. Recommended doses of NPK were 80, 40,
40 kg haÿ1, respectively. Nitrogen was given in form of
urea, phosphorus as single super phosphate and
potassium as murate of potash. Half dose of nitrogen
and full dose of phosphorus and potassium were given
as basal dressing and another half of nitrogen as top
dressing. After sowing, 32 pots were transferred to each
site. Pots were placed in unshaded open area receiving
uniform light. Micrometeorological variations in tem-
perature were 0.1–0.2  C, relative humidity 1–3%
between the sites. Light intensity was identical at all
sites. The pots were uniformly watered throughout the
experiment in order to maintain constant soil moisture.
For analysis, triplicate random samples of plants from
each treatment of each site were taken at 20 days after
sowing (DAS) and then at regular intervals of 15 days.
Final harvest was done on November 16, 2001 at 50
DAS. For total biomass determination, plants were oven
dried at 80  C until the constant weight was obtained
and values were expressed as g plantÿ1. The chlorophyll
content was expressed as mg gÿ1 dry leaf and measured
by using the method of Machlachlan and Zalik (1963).
Carotenoid content was calculated by the method of
Duxbury and Yentsch (1956). Protein analysis in fresh
leaves was performed by using the method of Lowry
et al. (1951). Ascorbic acid in fresh leaves was measured
using the 2,6 dichlorophenol indophenol method of
Keller and Schwager (1977). Catalase and peroxidase
enzyme activities were determined using the methods of
Kar and Mishra (1976) and Britton and Mehley (1955),
respectively. Determination of reducing and total soluble
sugars was performed by reference to glucose standards
using the calorimetric copper method of Somogyi (1952)
and for starch extraction, the method of McCready et al.
(1950) was followed.
Air monitoring of gaseous pollutants (SO2, NO2 and
O3) was done with the help of gas samplers kept at
30 cm height from ground at each site by using wet
chemical methods. SO2, NO2 and O3 were measured by
methods of West and Gaeke (1956), Merryman et al.
(1973) and Byers and Saltzman (1958), respectively. No
continuous advanced gas analyzers were available and
 A. Singh et al. / Environmental Pollution 134 (2005) 385–395 387

Fig. 1. Map of Allahabad city showing location of experimental sites.

gas samplers using wet chemical methods were the best
possible devices with the available resources. Monitor-
ing of pollutants was conducted for 6 h from 10 A.M. to
4 P.M. at weekly intervals at each site, this was the only
option to ensure the safety of the samplers and because
of frequent failure of electricity at various sites as
samplers have a battery back up of only 6 h.
Data were analyzed through three-way and two-way
ANOVA using SPSS software (SPSS Inc., version 10.0)
for assessing the significance of quantitative changes in
different parameters due to ambient air pollution.
3. Results
Results of air monitoring showed that RRP was the
most polluted sites among all experimental sites, where
SO2, NO2 and O3 were recorded in the range of 38.2–
65.0, 30.8–42.5 and 17.0–30.8 mg mÿ3, respectively. Min-
imum concentrations of SO2, NO2 and O3 ranged
between 10.6–18.3, 2.5–12.5 and 3.5–15.3 mg mÿ3, re-
spectively at site Ar (Table 3). Since all the pollutants
showed minimum concentrations at Ar, this site was
treated as reference site for comparing the levels of
changes in various parameters recorded at other sites
with relatively elevated levels of pollutants.
Total biomass of palak plants was reduced with
increasing pollution load at all sampling intervals
(Fig. 2) Significant negative correlations were found
between total biomass and SO2 (r Z ÿ0.92, p ! 0.01),
NO2 (r Z ÿ0.85, p ! 0.05) and O3 (r Z ÿ0.91,
p ! 0.01) (Table 4). F2 treatment showed a positive
response against air pollutants by increasing the total
biomass. Maximum total biomass (4.7 g plantÿ1) was
388 A. Singh et al. / Environmental Pollution 134 (2005) 385–395

Table 2 0.6
Brief description of experimental sites F0 F1 F2 F3 20 DAS

Total biomass (g plant-1)

0.5
Site code Experimental Character of site Distance (km)
site and direction 0.4
from city centre
AAI Allahabad Near bank of river 3 km south 0.3
Agricultural Yamuna, and national
0.2
Institute highway (NH-27); heavy
traffic, frequent congestion, 0.1
heavy vehicles, medium
density population. 0.0
Ar Arail Near bank of river 3 km south east 4.0 35 DAS

Total biomass (g plant-1)

Yamuna, open, small
population 3.5
Bh Bahrana City centre, near national 0 km
3.0
highway (NH-2), heavy
and light motor vehicles, 2.5
frequent traffic jams, high
2.0
density population.
Jh Jhunsi Near national highway 5 km east south 1.5
(NH-2) periurban area
Mh Mehdeori Near bank of river 5.5 km north 1.0
Ganga, light vehicles, 50 DAS

Total biomass (g plant-1)

4.5
periurban area.
CL Civil lines Commercial area, near 2.5 km
railway station, urban north west 4.0
area
RRP Rajrooppur Near national highway 6.5 km west 3.5
(NH-2), industries,
railway track, heavy 3.0
traffic, high density
population, urban area
2.5
AAI CL Mh Jh Bh Ar RRP
Experimental sites
recorded at site Ar in F2 treatment, which was 3.9 g
Fig. 2. Effect of air pollution on total biomass of palak plants grown at
plantÿ1 at F0 treatment. Total biomass was least at site different experimental sites with varying fertility levels.
RRP (3.3 g plantÿ1) under F0 treatment and it increased
to 4.3 g plantÿ1 due to F2 treatment (Fig. 2). Three-way
ANOVA test showed that the variations in total load (Fig. 3). Maximum total chlorophyll (0.95 mg gÿ1
biomass were significant ( p ! 0.001) due to plant age, dry leaf) and carotenoid contents (0.44 mg gÿ1 dry leaf)
site, nutrient treatment and their interactions (Table 5). were observed at Ar (reference site) and minimum at
Total chlorophyll and carotenoid contents in palak RRP, most polluted site (total chlorophyll 0.65 mg gÿ1
leaves were lower at sites experiencing higher pollution dry leaf; carotenoid 0.3 mg gÿ1 dry leaf) 50 DAS under
F2 treatment. Total chlorophyll and carotenoid contents
showed significant negative correlations with SO2
Table 3
Levels of SO2, NO2 and O3 at different sites during experiment (r Z ÿ1.0, p ! 0.001 and r Z ÿ0.99, p ! 0.001, respec-
(mg mÿ3) tively), NO2 (r Z ÿ0.96, p ! 0.001 and r Z ÿ0.98, p !
Gaseous Experimental sites 0.001, respectively) and O3 (r Z ÿ0.98, p ! 0.001 and
pollutants r Z ÿ0.99, p ! 0.001, respectively) (Table 4). ANOVA
CL Ar RRP Mh Jh Bh AAI
test showed that total chlorophyll and carotenoid
Sep., 2001
contents varied significantly due to plant age, site,
SO2 16.4 10.6 38.2 14.2 16.1 35.4 25.4
NO2 18.6 2.5 30.8 7.5 12.5 27.5 22.2 fertilizer treatment and their interactions except for
O3 10.2 3.5 17.0 6.5 8.4 16.4 12.5 plant age ! site ! fertilizer treatment interactions for
Oct., 2001
carotenoid (Table 5).
SO2 40.0 15.3 55.7 23.5 30.2 51.2 44.6 Protein and ascorbic acid contents showed significant
NO2 24.4 8.0 36.6 14.6 20.5 31.9 26.5 negative correlations with individual air pollutants
O3 14.3 10.3 27.6 10.0 11.7 22.5 18.6 (Table 4). Protein content increased with the increase
Nov., 2001 of plant age, while ascorbic acid decreased. Maximum
SO2 45.7 18.3 65.0 32.5 38.6 60.3 50.4 protein and ascorbic acid contents were observed at 50
NO2 27.5 12.5 42.5 18.7 23.6 37.5 31.6 and 35 DAS, respectively under F2 treatment at Ar
O3 20.4 15.3 30.8 15.9 16.9 26.4 24.0
(Fig. 4). Variations in for protein and ascorbic acid
 A. Singh et al. / Environmental Pollution 134 (2005) 385–395 389

contents were significant due to plant age, site, fertilizer

Reducing sugars
treatments (Table 5).
Catalase activity decreased with increasing levels of
ÿ0.99***
ÿ0.97***
ÿ0.99***

0.99***
0.99***
0.96***

0.98***
ÿ0.99***
0.97***
0.90**

ÿ0.91**
0.94*
air pollutants, while peroxidase activity increased

1.00
(Fig. 5). Catalase activity showed significant negative
correlations with SO2 (r Z ÿ0.97, p ! 0.001), NO2 (r Z
Soluble sugars

ÿ0.95, p ! 0.001) and O3 (r Z ÿ0.95, p ! 0.001), while

peroxidase activity showed highly significant positive
0.95***

ÿ0.95***
0.87**
ÿ0.88**
ÿ0.93**
ÿ0.90**

ÿ0.90**
0.94**
ÿ0.90**
0.82*

ÿ0.80*
correlation with SO2 (r Z 0.99, p ! 0.001), NO2 (r Z

1.00
0.95, p ! 0.001) and O3 (r Z 0.98, p ! 0.001) (Table 4).
Three-way ANOVA test showed significant variations in
ÿ0.96***

ÿ0.96***

0.98***
0.96***
0.97***
0.97***

ÿ0.98*** enzyme activities due to plant age, site and fertilizer

ÿ0.94**

0.85**

0.94*
Starch

1.00
treatment and their interactions except for plant
age ! site, site ! treatment and plant age ! site !
treatment interactions for catalase activity (Table 5).
Peroxidase activity

Starch and reducing sugar contents also decreased

with increasing levels of air pollutants while soluble
Correlation matrix of gaseous pollutants and total biomass and different physiological and biochemical characteristics of palak plants

sugars increased (Fig. 6). Correlation matrix showed

0.99***
0.95***
0.98***

ÿ0.99***
ÿ0.98***
ÿ0.96***
ÿ0.95***
ÿ0.94***
ÿ0.93**

a significant negative correlation between individual

1.00

pollutants and starch and reducing sugars and positive

correlation between soluble sugars and pollutants (Table
Catalase activity

4). Starch, reducing sugars and soluble sugars varied

significantly due to plant age, site, fertilizer treatment
ÿ0.97***
ÿ0.95***
ÿ0.95***

0.98***
0.98***

and their interactions except for plant age ! site,

0.94**
0.88**
0.81*

1.00

site ! treatment and plant age ! site ! treatment for

starch and soluble sugars (Table 5).
Ascorbic acid

ÿ0.95***
ÿ0.95***

4. Discussion
ÿ0.91**

0.94**
0.94**
0.91**
0.85*

1.00

In many cities of developing countries, the levels of

air pollutants often exceed toxic limits and adversely
ÿ0.96***

0.98***
ÿ0.89**
ÿ0.92**

0.94**
Protein

affect human health, vegetation and built cultural

0.85*

1.00

heritage. In urban areas of Allahabad city, high levels

of automobile emissions have elevated the levels of
Carotenoid

pollutants to an extent that inhibited the plant growth

ÿ0.99***
ÿ0.98***
ÿ0.99***

0.99***
0.89**

and reduced the yield of palak grown in urban and

1.00

periurban areas. Urban air quality of Varanasi, an

*p ! 0.05, **p ! 0.01, ***p ! 0.001 and NS Z not significant.

adjoining city of Allahabad has also been shown to

cause deleterious effects on woody transplants grown in
Total chlorophyll

urban areas (Pandey and Agrawal, 1994) and yield

losses in crop plants grown in periurban areas (Agrawal
ÿ0.99***
ÿ0.96***
ÿ0.98***
0.88**

et al., 2003).
1.00

Air monitoring conducted in Varanasi has shown

that SO2 concentration varied from 14 to 43 mg mÿ3,
NO2 from 16 to 34 mg mÿ3 and O3 from 12 to 42 mg mÿ3
Total biomass

during the rainy season (July–October) in urban areas

ÿ0.92**

ÿ0.91**

(Pandey et al., 1992). The levels of SO2 and NO2

ÿ0.85*

1.00

observed in the present study are similar to that of

Pandey et al. (1992), but O3 levels are low. From the
meteorological data it is clear that rains were frequent in
Peroxidase activity
Total chlorophyll

Catalase activity

Reducing sugars

September and October, and hence all the pollutants

Soluble sugars
Total biomass

Ascorbic acid

including O3 showed lower values during these months.

Carotenoid

In November, however, O3 formation increased with

Table 4

Protein

Starch

a longer sunshine period. The permissible annual safe

NO2
SO2

O3

limits set by CPCB, India for 8 hourly SO2 and NO2

390 A. Singh et al. / Environmental Pollution 134 (2005) 385–395

Table 5
Variance ratio for total biomass, photosynthetic pigments, protein, ascorbic acid, enzyme activities and carbohydrate content of palak plants grown
with different fertility levels at various experimental sites
Parameter Plant age (A) Site (B) Treatment (C) A!B A!C B!C A!B!C
Total biomass *** *** *** *** *** *** ***
Total chlorophyll *** *** *** *** *** *** ***
Carotenoid *** *** *** *** *** *** NS
Protein *** *** *** NS *** NS NS
Ascorbic acid *** *** *** NS *** NS NS
Catalase activity *** *** *** NS *** NS NS
Peroxidase activity *** *** *** * *** ** **
Starch *** *** *** NS *** NS NS
Soluble sugars *** *** *** NS *** NS NS
Reducing sugars *** *** *** ** *** *** ***
*p ! 0.05, **p ! 0.01, ***p ! 0.001 and NS Z not significant.

concentrations in urban areas are 60 mg mÿ3. SO2 permissible safe limit set for O3 in India. Since the
concentration crossed this limit at two sites (RRP and monitoring of pollutants was conducted on 6 hourly
Bh) during September. NO2 concentration remained sample collections, the measurement of peak concen-
always below the permissible limit. There is no trations in between cannot be provided. In Varanasi, 2 h

0.7
F0 F1 20 DAS F0 F1 20 DAS 0.4
0.6 F2 F3 F3
F2

0.5 0.3

0.4
0.2
0.3
0.1
0.2

0.1 0.0
0.5
Total chlorophyll (mg g-1 dry leaf)

0.8 35 DAS 35 DAS

Carotenoid (mg g-1 dry leaf)

0.4
0.7

0.6 0.3

0.5
0.2
0.4
0.1
0.3

0.2 0.0
1.0 50 DAS 50 DAS 0.5

0.4
0.8

0.3
0.6
0.2

0.4
0.1

0.2 0.0
AAI CL Mh Jh Bh Ar RRP AAI CL Mh Jh Bh Ar RRP
Experimental sites Experimental sites

Fig. 3. Effect of air pollution on total chlorophyll and carotenoid contents in leaves of palak plants grown at different experimental sites with varying
fertility levels.
 A. Singh et al. / Environmental Pollution 134 (2005) 385–395 391

14 15 400
F0 F1 F0 F1 F2 F3
13

Protein (mg g-1 fresh leaf)

Protein (mg g-1 fresh leaf)

F2 F3
14

Starch (mg g-1 dry leaf)

12 375
11 13
10 350
9 12

8 325
11
7 35 DAS 50 DAS
6 10
300
Ascorbic acid (mg g-1 fresh leaf)

Ascorbic acid (mg g-1 fresh leaf)

0.90

Soluble sugars (mg g-1 dry leaf)

1.6
300

0.75 275

1.2 250

0.60
225

50 DAS 200
35 DAS
0.8 0.45
AAI CL Mh Jh Bh Ar RRP AAI CL Mh Jh Bh Ar RRP 175

Reducing sugars (mg g-1 dry leaf)

Experimental sites Experimental sites

Fig. 4. Effect of air pollution on protein and ascorbic acid contents in

leaves of palak plants with varying fertility levels at different 250
experimental sites.

200

90 150
(mM H2O2 decomposed min-1 g-1 fresh leaf)

AAI CL Mh Jh Bh Ar RRP
F0 F1
80 Experimental sites
F2 F3
70
Fig. 6. Effect of air pollution on starch, soluble and reducing sugar
Catalase activity

contents in leaves of palak plant at 50 DAS sites with varying fertility

60 levels at different experimental.

50 peak concentrations of SO2 varied from 25 to 95 mg mÿ3,

NO2 from 27 to 61 mg mÿ3 and O3 from 21 to 102 mg mÿ3
40
in periurban and urban areas (Pandey et al., 1992). Peak
30 concentration affects vegetation more adversely than
prolonged exposure to low concentrations (Lefohn and
20
Jones, 1986). The lower O3 concentration recorded
5 during the present study may also be ascribed to the
(mM purpurogallin formed min-1 g-1 fresh leaf)

presence of monitoring sites near roads, where O3 is

quickly scavenged.
4
The adverse effects of urban air pollutants are clearly
evident on physiological and biochemical processes of
Peroxidase activity

3 palak plants during the present investigation. Photo-

synthetic pigments are fairly sensitive to air pollutants
2 and their sensitivity may determine the responses of
plants to pollutants. Significant negative correlations
1 were obtained between air pollutants and total chloro-
phyll and carotenoid contents. Khan and Khan (1994)
0
reported that combined treatments of O3 and SO2 at all
AAI CL Mh Jh Bh Ar RRP concentrations had a significant negative effect on the
Experimental sites leaf pigments. Exposure to SO2 causes more reduction in
Fig. 5. Effect of air pollution on catalase and peroxidase activity in
chlorophyll than carotenoids in wheat plants (Verma
leaves of palak plants at 50 DAS with varying fertility levels at and Agrawal, 2001). El-Khatib (2003) also reported
different experimental sites. reductions in chlorophyll and carotenoid contents
392 A. Singh et al. / Environmental Pollution 134 (2005) 385–395
caused by elevated levels of O3 in five common Egyptian
plant species. Nutrient amendment in various combina-
tions significantly lowered the magnitude of reduction in
chlorophyll as compared to unamended plants (F0
treatment). Verma et al. (2000) also reported that
nutrient amendment lowered the magnitude of reduc-
tion in chlorophyll content as compared to unamended
ones, confirming the results of the present investigation.
Nitrogen supply is reported to increase the leaf
photosynthesis via the amount of N-containing compo-
nents such as ribulose-1,5 bisphosphate carboxylase/
oxygenase activity (Sivasankar et al., 1993) and also by
chlorophyll formation (Agrawal and Verma, 1997).
However, N and P deficiency reduces the chlorophyll
concentration (Rousseau and Reid, 1990).
Air pollutants are known to induce the degradation of
biologically important molecules such as proteins with
the consequent release of malondialdehyde (Mudd,
1982). Protein content showed significant negative cor-
relations with air pollutants and significant increase in
protein content was observed due to application of
mineral nutrients at all sites. The protein content
depends upon N uptake and plants receiving a higher
dose assimilate more N as compared to the unamended
plants. Plants grown with higher N-supply invest
a greater proportion of carbon in protein (Makino et al.,
1984). Verma and Agrawal (1996) also noticed that N, P
and K amendment in soybean plants significantly
reduced the levels of decrease in protein content of SO2
exposed plants.
Ascorbate is a ubiquitous soluble antioxidant in
photosynthetic organisms and the most important
reducing substrate for H2O2 detoxification. It has been
suggested that pollutants produce oxyradicals in plants
(Shimazaki et al., 1980; Sakaki et al., 1983). These
radicals cause widespread damage to membranes and
associated molecules including the chlorophyll pigments
(Sakaki et al., 1983). Several authors have reported that
ascorbic acid can serve as a free radical scavenger
against O3 (van Hove et al., 2001; El-Khatib, 2003). The
reduction in ascorbic acid concentration may be
ascribed to its consumption during removal of cytotoxic
free radicals generated in chain reactions after the
penetration of oxidative pollutants into leaf tissues.
Agrawal and Verma (1997) also observed higher
ascorbic acid content in SO2 exposed plants amended
with fertilizers.
Peroxidase activity showed an increasing trend with
increasing pollution levels at various sites, while catalase
activity decreased. Singh (1998) also found that wheat
plants exposed to O3 showed increase in peroxidase
activity without any specific symptoms of O3 on foliage.
Tingey et al. (1975) stated that stimulation of peroxidase
activity in pollutant-exposed plants might be due to
increased oxidative processes under pollutant stress.
Present observations are in conformity with the earlier
findings of Ranieri et al. (1997), who showed that
catalase activity decreased with increase in SO2 levels
while peroxidase activity increased.
Starch content showed significant negative correla-
tion with individual pollutants. Rennenberg et al. (1996)
suggested that O3 probably interacts with carbon
allocation by inhibiting sucrose export. This causes an
accumulation of starch in leaves, which results in
reduction of photosynthesis and consequently reduces
the level of starch in plants. Nutrient amendment in
different combinations has significantly elevated the
levels of starch in plants. Agrawal and Verma (1997)
also reported reduction in foliar starch content of two
cultivars of wheat treated with SO2 compared with
untreated plants and also found significant increase after
nutrients application.
An increase in total soluble sugars was recorded
with increasing pollution levels while reducing sugars
reduced. The adverse effects of air pollutants are evident
in the form of changes in pool volume of free carbo-
hydrates in palak grown at different sites with various
fertility levels. Katase et al. (1983) reported that SO2-
induced inhibition of photosynthesis in rice reduced the
level of starch in the plants. An increase in concentration
of sugar was associated with reduced starch content,
suggesting increased hydrolysis of polysaccharides into
monosaccharides due to gaseous pollution (Koziol and
Jorden, 1978). Nutrient amendment has significantly
reduced the level of decrease in starch and reducing sugar
contents at various sites, which might be due to increased
photosynthetic rate led by nutrient amendment. Meyer
et al. (2000) reported that O3 caused inhibition of
photosynthesis, and consequently decline in assimilate
production. Agrawal et al. (2003) have also reported
significant reductions in photosynthetic rate of a number
plant species growing in a periurban area of Varanasi
experiencing higher level of pollutants.
Total biomass accumulation reduced in palak with
increasing pollution load at various sites. This suggests
that air pollutants directly interfere with various
fundamental processes of plants, resulting in lower
biomass accumulation. Ashmore et al. (1987) have also
reported a decline in biomass accumulation in different
plant parts along a gradient of air pollution around
London. Agrawal et al. (2003) have also reported a
negative correlation between ambient air pollutant levels
and biomass accumulation in plants grown in the out-
skirts of Varanasi city experiencing similar climatic
conditions. Verma et al. (2000) reported that 390 mg mÿ3
SO2 treatment for 4 h daily for 5 days weekÿ1 for 8
weeks resulted in a significant reduction in biomass
accumulation and productivity in wheat plants were due
to the integrated result of effects on a range of bio-
chemical, physiological and metabolic activities in
plants. The joint action of O3 and SO2 caused significant
suppression in dry matter of tomato shoot and root at
 A. Singh et al. / Environmental Pollution 134 (2005) 385–395
all concentrations (Khan and Khan, 1994). McKee et al.
(1997) also reported that elevated O3 caused a 15%
decline in total biomass accumulation in wheat plants.
Fertilizer amendment has a significant effect on plant
response to air pollutants. Agrawal and Verma (1997)
reported that plant height and total biomass reduced
significantly in SO2 treated plants, except those grown
using recommended and twice recommended N, P and
K applications. In the present investigation, F2 treat-
ment showed the most positive impact on biomass
accumulation and photosynthetic pigments by decreas-
ing the negative impact of air pollutants. The percent
reduction in total biomass at 50 DAS was 15.5 and
9.5%, respectively at RRP site in nutrient unamended
and amended plants as compared to the same growing at
Ar site experiencing lowest levels of air pollutants. The
double recommended dose of nutrients showed maxi-
mum percent reduction in total biomass (29% at RRP
site compared to Ar site) suggesting that this dose is
supra optimal and caused negative influence on the
plants. Application of nutrients higher than the demand
has been shown to reduce the positive effects of
fertilizers (Agrawal and Verma, 1997; Verma et al.,
2000). Recently, Singh et al. (2003) also revealed that air
pollutants suppressed the growth and yield of wheat
plants grown at various urban and periurban sites of
Allahabad but fertilizer amendment higher than the
recommended dose resulted in a positive response by
increasing the total biomass, weight of 1000 seeds and
yield.
The supply of macro nutrients N, P and K increased
the total biomass in palak by increasing the levels of
photosynthetic pigments, antioxidative property and
metabolites in foliar tissue, which have further reduced
the magnitude of reduction in biomass due to air
pollutants compared to unfertilized plants. Coleman
et al. (1989) have suggested that plants growing in
nutrient poor conditions may be more sensitive to air
pollution with respect to changes in carbon gain. N
limitation has been shown to decrease chlorophyll and
protein contents, RuBP carboxylase activity and in-
crease the mesophyll resistance, which all limit CO2
fixation (Osman and Milthorpe, 1971). High P avail-
ability is found to increase the rate of photosynthesis
(Rousseau and Reid, 1990). K fertilization is also
beneficial due to its role in stomatal opening, photosyn-
thesis, protein synthesis and osmotic and pH regulation
(Wyn Jones and Pollard, 1983).
5. Conclusions
The data obtained in the present investigation suggest
that both air pollutants and nutrient deficiency have
caused adverse impact on various physiological and
biochemical processes and total biomass accumulation
393
of palak plants grown at sites experiencing elevated
pollutant concentrations. Increasing pollution load also
deteriorated the nutritive quality of palak plants, as
protein and carbohydrate contents were decreased. One
and half times of recommended dose of NPK was most
efficient in reducing the adverse effects of air pollutants
on palak plants. The present investigation also suggests
that urban air quality of Allahabad city is unfavourable
for vegetable production in urban and periurban areas.
Though the concentrations of individual pollutants were
not very high except SO2, the levels of reductions were
fairly significant. This clearly shows that pollutants in
combination may have acted synergistically in causing
greater adverse impact. Low concentrations of pollu-
tants have been shown to increase the stomatal
conductance thus facilitating the pollutant uptake and
consequently greater negative response. Palak plants
seem to be fairly sensitive to air pollutants under
ambient conditions. More large-scale studies are,
however, required to ascertain the potential for use of
this plant as a biomonitor of air pollution.
Acknowledgements
Authors wish to express sincere thanks to Prof. R.B.
Lal, Vice Chancellor, Allahabad Agricultural Institute -
DU and Prof. P.W. Ramteke, Director (Research) for
providing laboratory facilities and encouragements and
to C.S.I.R. (New Delhi) for providing financial support.
Authors are also grateful to Professor Madhoolika
Agrawal (B.H.U.) and to anonymous reviewers for
comments and fruitful suggestions.
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