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Israel Journal of Plant Sciences
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Soil amendments of fly ash: effects on function and
biochemical activity of Carthamus tinctorius L. plants
a

b

a

a

a

Qurratul , Sumira Jan , Riyazzuddin Khan , Mahmooduzzafar & T.O. Siddiqi
a

Department of Botany, Jamia Hamdard, New Delhi-62, India

b

Centre of Research for Development (CORD), Kashmir University, India
Published online: 02 Oct 2014.

To cite this article: Qurratul, Sumira Jan, Riyazzuddin Khan, Mahmooduzzafar & T.O. Siddiqi (2014): Soil amendments of fly
ash: effects on function and biochemical activity of Carthamus tinctorius L. plants, Israel Journal of Plant Sciences, DOI:
10.1080/07929978.2014.945313
To link to this article: http://dx.doi.org/10.1080/07929978.2014.945313

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Israel Journal of Plant Sciences, 2014
http://dx.doi.org/10.1080/07929978.2014.945313

Soil amendments of fly ash: effects on function and biochemical activity
of Carthamus tinctorius L. plants
Qurratula, Sumira Janb*, Riyazzuddin Khana, Mahmooduzzafara and T.O. Siddiqia
a

Department of Botany, Jamia Hamdard, New Delhi-62, India; bCentre of Research for Development (CORD),
Kashmir University, India

Downloaded by [The University of Kashmir ], [Sumira Jan] at 00:39 13 October 2014

(Received 2 June 2014; accepted 7 July 2014)
Proper disposal and recycling of different industrial waste materials have long been recognized as a prime environmental
concern. The present study evaluated the effects of soil amendment of fly ash, a major industrial waste material, on soil
properties, plant growth, productivity and metabolites production of safflower (Carthamus tinctorius L.). The soil was
amended with varied concentrations of fly ash (0%, 5%, 10%, 25%, 50%, 75% per pot) prior to sowing under field
conditions in the herbal garden of Jamia Hamdard. Sampling was conducted at different growth stages, i.e. pre-flowering,
flowering and post-flowering. Our results demonstrate that fly ash concentrations up to 25% improved the physicochemical
properties of the soil as compared to non-treated control resulting in increased availability to the plant of macro and
micronutrients and thereby stimulating plant growth and productivity. Contents of photosynthetic pigments, sugars,
protein, and nitrate reductase (NR) activity increased under 25% fly ash amendment. The highest beneficial effect was
found during the flowering stage > pre-flowering > post-flowering stages. Further increase in fly ash concentration
reduced the stimulated effects on the plants, exhibiting a minimum under 75% fly ash application. Thus, it was concluded
that incorporation of 25% fly ash to the cultivation soil not only improves the physicochemical properties of the soil, but
also contributes to better growth, yield and metabolism of safflower.
Keywords: fly ash; Carthamus tinctorius (safflower); growth attributes; photosynthetic pigments; metabolic contents

Introduction
Fly ash is formed of minute glossy particles ranging in
size from 0.01 to 100 mm that are the combustion byproduct of coal (Davison et al. 1974). Indian thermal
power plants produce more than 100 million tons of fly
ash per annum, which is expected to reach 175 million
tons in the near future (Jamwal 2003). Fly ash production
depends on the quality of coal, which contains a relatively
high proportion of ash that leads to 10
30% of its formation (Singh et al. 2008). According to World Bank, India
will require 1000 km2 of land for the disposal of nearly
65 million of coal ash till 2015 (Parisara 2007). Therefore,
there is a need for new and innovative methods to reduce
the potential negative impacts of fly ash on the environment. Further, it is necessary to develop possible avenues
for its disposal. One possibility is the utilization of fly ash
as a soil amendment to facilitate efficient plant growth.
Fly ash is alkaline associated with small quantities of Fe,
Ca, Mg, Na, K, Ti and P oxides (Aitken et al. 1984). It
contains several nutrients which are beneficial for plant
growth, as well as toxic heavy metals such as Cr, Pb, Hg,
Ni, V, As and Ba (Robab et al. 2010). Addition of fly ash
to soil neutralizes the acidity to a level suitable for agricultural crops and increases the availability of Na, K, Ca,
*Corresponding author. Email: [email protected]
Ó 2014 Taylor & Francis

Mg, B, SO4¡2 and other nutrients except N (Sarangai
et al. 2001). The higher dose of these toxic metals and
metalloids leads to alteration of enzyme activities, degradation of membranes, proteins, nucleic acids and chlorophyll, and increased per-oxidation of lipids due to
generation of free radicals (Gupta et al. 2007); and at
lower concentration could enhance the growth and yield
of many crop plants. Foliar application of fly ash
increased plant height, metabolic rate and photosynthetic
pigment content in Zea mays and Glycine max (Mishra &
Shukla 1986). Fly ash amendment is also reported to
improve the performance of oil-seed crops such as sunflower (Helianthus sp.), Sesame (Sesamum indicum),
turnip (Brassica rapa) and groundnut (Arachis hypogaea)
(Basu et al. 2009). There are also reports that 10
25% utilization of fly ash resulted in better growth, dry matter
production, and increased photosynthetic pigments in
Lactuca sativa and Indian rice cultivars (Srivastava et al.
1995; Dwivedi et al. 2007).
Carthamus tinctorius L., commonly known as safflower, is an intriguing plant species ranging from the
western borders of China to the Mediterranean sea (Harrathi et al. 2012). At present, safflower is grown in more
than 30 countries. India, Mexico and USA contribute

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2

Qurratul et al.

approximately 70% of world safflower oil production
(Sehgal et al. 2009). It is a multipurpose crop grown for
its foliage, flowers and seeds which have numerous foodrelated biological properties and multiple functional uses
(Bowles et al. 2010). The linolenic acid-rich oil of safflower is known to reduce blood cholesterol level and is
used clinically for the treatment of cataclasis, osteoporosis
and rheumatoid arthritis (Lee et al. 2002). In traditional
Chinese medicine, the flowers were used for the treatment
of cardiovascular and cerebral troubles (Gao et al. 2000).
More recently, it has been reported that flower extracts
have antioxidants, antibacterial, anti-inflammatory, antidepressant and antitumor properties (Hiramatsu et al.
2009). As far as its nutritional value is concerned, safflower seeds are widely used for the preparation of margarine and it is marketed as edible oil in Germany and USA.
The flowers are the source of natural orange, yellow and
red dye, which is used as alternative to saffron (Crocus
sativus). The fruits are widely used as birdseeds and often
incorporated in livestock feed (Rammal et al. 2009). The
importance of safflower has increased in recent years,
especially with interest in the production of bio fuels
(Dordas & Sioulas 2008). Safflower is illustrated to be
more drought-resistant and salt-tolerant than some other
oil crops (Bassil & Kaffka 2002). There has been no
report on the effect of fly ash on Carthamus tinctorius L.
The present study is therefore an effort to investigate the
impact of fly ash amendment on the growth, biomass,
yield and some biochemical responses of Carthamus
tinctorius L.

under natural conditions in 12-inch pots filled with 8 kg of
varying soil and fly ash mixtures per pot. The fly ash and
soil were mixed together in six proportions, i.e. 0, 5, 10,
25, 50, and 75% (w/w), and designated as T0, T1, T2, T3,
T4 and T5, respectively.

Materials and methods

Physicochemical analysis of fly ash amended soil

Source of fly ash

Fly ash treatment

Soil pH was determined in a 1:2 soil: solution suspension
using a glass electrode digital pH meter (Jackson 1968).
Electrical conductivity (EC) was determined with a digital
conductivity meter in the supernatant of the soil water suspension (1:2 ratio) (Jackson 1968). Water holding capacity (WHC) and bulk density (BD) were determined by the
method of Black (1965). Organic carbon (OC) in the soil
was estimated by the modified rapid titration method of
Walkley and Black (Allison 1973). Soil-available phosphorus (P) was determined following Olson et al. (1954).
Soil-available potassium (K) was extracted with neutral N
ammonium acetate and determined using a flame photometer (Hanway & Heidel 1952). Nitrogen (N) and sulphur
(S) content (% dry weight) were analyzed by packing a
known weight of fly ash amended soil sample powder in
tin boats with the help of Elementar system (CHNS
Analyse, Vario EL).

The experiment was carried out at the herbal garden of
Jamia Hamdard, New Delhi, India (28 380 N, 77 110 E; elevation of 228 m), about 70
80% relative humidity, 35  C
temperature. The fly ash and normal soil were both dried
under sunlight for 7 days. The experiment was conducted

Morphological parameters
Plant height was measured in centimeters, number of
branches and leaves per plant were counted at three

Fly ash used in the experiment was collected from
the Badarpur Thermal Power Plant, located at the
Badarpur border on Delhi
Mathura Road (latitude
28 290 51.3300
28 300 41.4100 N; longitude 77 180 09.6300

77 180 40.0100 E; elevation 211 m a.m.s.l.). Badarpur Thermal Power Station comprises 3 units of 100 MW each and
2 units of 210 MW each and has a power generating
capacity of 720 MW on consumption of about 10,000
tonnes of low-grade bituminous coal daily. It emits 1450
tonnes fly ash and 600 tonnes SO2 per day. The soil at the
experimental field was sandy loam in texture, with pH
7.38 and an electrical conductivity of 0.235 ds/m.

Plant material
The crop plant selected for study was safflower (Carthamus tinctorius L.). It is a tap-root herbaceous, annual oilseed plant belonging to the family Asteraceae. The seeds
of safflower were procured from the Medico Botany Laboratory, Department of Botany, Faculty of Science, Jamia
Hamdard, New Delhi.
Experimental setup
The field experiment was repeated three times, each with
five replicates per treatment. Seeds were sown in the first
week of October 2011. Other agricultural practices such
as irrigation and weeding were carried out as is customary
in the region. The planted seeds were observed daily until
germination commenced. The dates of commencement
and termination of germination and the number of seeds
that germinated each day were recorded. The seedlings
were collected at three phenological stages: pre-flowering
(45 days after sowing, DAS), flowering (90 DAS) and
post-flowering (135 DAS). Leaves were collected for the
analyses of growth, yield attributes and photosynthetic
pigments at the physiological maturity of the plant.

Israel Journal of Plant Sciences
developmental stages, i.e. pre-flowering, flowering and
post-flowering stages.
For plant biomass measurement, each plant was separated into leaves, roots and stems. The plant parts were
dried at 55  C for 48 h in Precision TM mechanical convection oven (GCA Corporation, San Francisco, USA)
and weighed with a Mettler TM balance (type H6,
§ 0.1 mg). Each plant part, including the root, was
weighed separately.

3

total chlorophyll and carotenoid contents using the equations given by Maclachlan and Zalik (1963) for chlorophyll a, Duxbury and Yentsch (1956) for chlorophyll b,
Arnon (1949) for total chlorophyll and Barnes et al.
(1992) for carotenoid contents.
Soluble protein content. Soluble protein content was
analyzed following the method of Bradford (1976).
Nitrate reductase activity. Nitrate reductase activity was
estimated according to Klepper et al. (1971).

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Yield characteristics
Soluble sugar content. The soluble sugar was estimated
by method of Dey (1990).

Number of heads per plant, number of seeds per head,
weight of 100 seeds per head and weight of seed per head
were determined at the physiological maturity stage.
Weights were recorded with a digital balance (Mettler
TM type H6, § 0.1 mg).

Statistical analysis
Each experiment was repeated three times with five replicates. The data are expressed as mean § SE (n D 5). Differences between several mean values were evaluated by
analysis of variance (ANOVA). Duncan’s multiple range
test (DMRT) at p < 0.05 level was used to determine
whether the values were significantly different from the
control. All the statistical tests were performed using
SPSS software (SPSS Inc., version 16.0).

Biochemical parameters
Photosynthetic pigment. Chlorophyll and carotenoid
contents were measured following Hiscox and Israelstam
(1979). Leaves, kept on a moist filter paper in an icebox,
were washed with cold distilled water. Leaf discs were
taken from either side of the midrib at the intraveinal
region for the determination of chlorophyll and carotenoid
content. One hundred milligrams of the chopped leaf
material were placed in vials containing 7 ml of dimethyl
sulfoxide (DMSO), in five replicates. The vials were kept
in an oven at 65 C for 1 h for complete leaching of the
pigments. Thereafter, the volume was brought up to 10 ml
with DMSO. The chlorophyll content was measured
immediately as absorbance at 480, 510, 645 and 663 nm
(Beckman spectrophotometer, model DU 640, Fullerton,
USA) against a DMSO blank. Values of optical densities
(ODs) were used to compute chlorophyll a, chlorophyll b,

Results
Physicochemical properties of fly ash and soil
before sowing
The physicochemical property of the soil and the fly ash
samples carried out in the present study is given in Table 1.
Soil pH significantly (p < 0.05) increased with application
of fly ash (from 7.38 in the control (T0) to 7.86 in fly ash
amended soil (T5)). EC value increased (p < 0.05) with
increasing concentration of fly ash application from 5% to

Table 1. Selected physicochemical properties of fly ash amended soils used in the study before sowing.
Fly ash amendments % (w/w)
T0 control
Parameters
pH (1:2, soil: water)
EC (dS/m)
WHC (%)
BD (g/cm3)
OC%
Available P (mg/kg)
Available K (g/kg)
N (%)
S (%)

T1

0%FA

T2

5%FA

7.38 § 0.001
0.24 § 0.001f
40.37 § 0.24f
1.58 § 0.001a
0.43 § 0.002f
172.0 § 0.32f
1.57 § 0.002f
0.11 § 0.001a
0.09 § 0.002f
f

T3

10%FA

7.52 § 0.001
0.29 § 0.002e
43.10 § 0.28e
1.52 § 0.001b
0.45 § 0.001e
196.68 § 0.27e
1.75 § 0.004e
0.09 § 0.001b
0.10 § 0.002e
e

T4

25%FA

7.65 § 0.001
0.89 § 0.002d
46.64 § 0.25d
1.42 § 0.001c
0.47 § 0.001d
204.77 § 0.30d
2.36 § 0.018d
0.08 § 0.001c
0.12 § 0.001d
d

T5

50%FA

7.68 § 0.002
0.92 § 0.010c
51.62 § 0.42c
1.41 § 0.001d
0.51 § 0.001c
236.56 § 0.22c
2.52 § 0.005c
0.07 § 0.001d
0.15 § 0.001c
c

75%FA

7.72 § 0.01
1.10 § 0.012b
54.89 § 0.33b
1.38 § 0.001e
0.53 § 0.002b
251.97 § 0.35b
2.67 § 0.002b
0.06 § 0.001e
0.27 § 0.002b
b

7.86 § 0.02a
1.23 § 0.013a
59.54 § 0.315a
1.35 § 0.001f
0.57 § 0.002a
289.90 § 0.38a
2.73 § 0.003a
0.05 § 0.001f
0.36 § 0.002a

Values represent mean § SE (n D 5). Values with different superscripts are significantly (p < 0.05) different from each other by DMRT (Duncan’s multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash (FA).

Values in parentheses indicate percent variation with reference to respective controls. Values represent mean § SE (n D 5). Values with different superscripts are significantly (p < 0.05) different from each
other DMRT (Duncan’s multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash (FA).

0.23 § 0.002f (28.12)
2.55 § 0.001f (32.18)
5.30 § 0.008f (23.63)
0.27 § 0.003e (15.62)
3.11 § 0.001e (17.28)
5.89 § 0.074e (15.12)
0.68 § 0.001a (112.50)
7.11 § 0.003a (89.09)
12.22 § 0.011a (76.08)
0.36 § 0.002c (12.5)
4.78 § 0.005c (27.12)
7.52 § 0.021c (8.36)
(C) Shoot dry weight (g)
Pre-flowering
0.32 § 0.003d
Flowering
3.76 § 0.003d
Post-flowering
6.94 § 0.003d

0.44 § 0.002b (37.50)
5.85 § 0.003b (55.58)
8.95 § 0.010b (28.96)

0.65 § 0.001f (31.57)
4.66 § 0.002f (22.3)
9.43 § 0.002f (28.01)
0.78 § 0.001e (17.89)
5.61 § 0.24e (6.5)
11.14 § 0.004e (14.96)
2.49 § 0.002a (162.10)
13.45 § 0.002a (124.16)
21.27 § 0.02a (62.36)
1.10 § 0.001c (15.78)
8.8 § 0.003c (46.66)
16.33 § 0.09c (24.65)
(B) Shoot fresh weight (g)
Pre-flowering
0.95 § 0.001d
Flowering
6.0 § 0.006d
Post-flowering
13.10 § 0.03d

1.58 § 0.001b (66.31)
10.39 § 0.001b (73.16)
18.40 § 0.03b (40.45)

5.34 § 0.07f (36.12)
48.4 § 0.24f (7.63)
55.8 § 0.2f (10.86)
6.12 § 0.12e (26.79)
50.4 § 0.24e (3.81)
59.6 § 0.24e (4.79)
14.4 § 0.17a (72.24)
73 § 0.16a (39.31)
82.32 § 0.19a (31.50)
11.9 § 0.1b (42.34)
65.7 § 0.37b (25.38)
74 § 0.47b (18.21)
9.7 § 0.22c (16.02)
61.2 § 0.2c (16.79)
70.56 § 0.23c (12.71)
(A) Shoot length (cm)
Pre-flowering
8.36 § 0.10d
Flowering
52.4 § 0.24d
Post-flowering
62.6 § 0.19d

75%FA
50%FA
25%FA
5%FA
0%FA

10%FA

T4
T3
T2
T1
T0 control

Morphological parameters
Shoot height, fresh and dry biomass of Carthamus tinctorius L. showed significant (p < 0.05) variation at all stages
of plant development (Table 2A
C). Shoot length, shoot
fresh and dry biomass were increased progressively from
5% to 25% fly ash applications as compared to control.
The highest increase was recorded in T3, with 25% of fly
ash supplementation at all stages of plant development.
With further increase in percentage of fly ash (i.e. in T4
and T5) the morphological development of the plants was
decreased and the lowest values were found at the 75% fly
ash treatment. However, the maximum increase (72.24%)
in shoot length was recorded in T3 plant at the pre-flowering stage, followed by the flowering (39.31%) and the
post-flowering (31.50%) stages. While in the case of shoot
fresh weight, maximum increase (163.53%) was recorded
in T3 at pre-flowering stage followed by flowering
(124.16%) and post-flowering (62.30%) stages. Shoot dry
weight followed the same trend as shoot fresh weight.
Root length, fresh weight and dry weight of Carthamus tinctorius L. showed significant increase (p < 0.05)
in T2 and T3 with 10 and 25% fly ash mixture at all stages
of plant development compared to control (Table 3A
C).
The highest increase was recorded in T3 (25% of fly ash)
and decreased under higher application rates. Moreover,
the maximum increase in root length (57.68%) was
recorded with T3 at post-flowering stage, followed by
flowering (55.49%) and pre-flowering (52.76%) stages.
While, in the case of root fresh weight, the maximum
increase (152.02%) was recorded in T3 at the flowering
stage, followed by post-flowering (107.3%) and pre-flowering (68.76%) stages. The same trend was observed in
root dry weight.
The number of branches showed a significant (p <
0.05) increase from 5% to 25% fly ash treatments at all
stages of plant development (Figure 1A). The highest

Parameters/stages

75%. The maximum EC (1.23 dS/m) was found in T5 with
75% fly ash level while in the control, it was 0.24 dS/m.
The same trend was noticed in WHC, demonstrating a
considerable increase of 6.50%, 15.53%, 27.90%, 35.96%
and 47.48% at 5%, 10%, 25%, 50% and 75%, respectively. The bulk density of the soil without fly ash addition
(T0) was the highest at 1.58 g/cm3, which significantly
(p < 0.05) decreased up to 1.353 g/cm3 in T5. As compare
to normal soil, organic carbon content increased under the
fly ash amended soil. Available P, K and S contents
increased significantly with fly ash. T5 had the highest P
(289.90 mg/kg), K (2.73 g/kg) and S (0.36%), and the
lowest levels were found in T0 (P D 172 mg/kg, K D
1.57 g/kg and S D 0.09%). Nitrogen (%) significantly
decreased with increased level of fly ash with the lowest
level found in T5 (0.05%), and the highest in the control
(0.11%).

T5

Qurratul et al.

Table 2. Effect of fly ash on shoot length (cm/plant), shoot fresh weight (g/plant) and dry weight (g/plant) at various growth stages of Carthamus tinctorius L.

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4

0.77 § 0.005b (21.45)
2.01 § 0.004b (35.81)
5.42 § 0.055b (72.06)
0.123 § 001b (23)
1.19 § 0.002b (158.6)
3.00 § 0.002b (80.72)

0.67 § 0.003c (5.67)
1.63 § 0.003c (10.13)
4.58 § 0.006c (45.39)
0.12 § 0.001c (20)
0.68 § 0.001c (47.82)
2.35 § 0.001c (41.56)

(B) Root fresh weight (g)
Pre-flowering
0.634 § 0.004d
Flowering
1.48 § 0.002d
Post-flowering
3.15 § 0.003d

(C) Root dry weight (g)
Pre-flowering
0.1 § 0.001d
Flowering
0.46 § 0.001d
Post-flowering
1.66 § 0.001d
0.15 § 0.001a (50)
1.74 § 0.004a (278.26)
3.14 § 0.001a (89.15)

1.07 § 0.002a (68.76)
3.73 § 0.104a (152.02)
6.53 § 0.007a (107.3)

9.96 § 0.0.04a (52.76)
22.64 § 0.112a (55.49)
27.28 § 0.620a (57.68)

25%FA

T3

0.072 § 0.001e (28)
0.39 § 0.036e (15.21)
1.17 § 0.04e (29.46)

0.52 § 0.002e (17.98)
1.2 § 0.003e (18.91)
3.01 § 0.018e (4.44)

5.96 § 0.067e (8.58)
11.8 § 0.2e (18.95)
15.4 § 0.187e (10.98)

50%FA

T4

0.06 § 0.001f (40)
0.301 § 0.001f (34.56)
0.99 § 0.001f (40.398)

0.38 § 0.003f (40.06)
0.8 § 0.003f (45.94)
2.07 § 0.011f (34.28)

4.96 § 0.050f (23.92)
8.02 § 0.215f (44.91)
13.1 § 0.10f (24.27)

75%FA

T5

Number of leaves/ plant

Number of branches/ plant

Values in parenthesis indicates percent variation with reference to respective controls. Values represent mean § SE (n D5). Values with different superscripts are significantly (p < 0.05) different from each
other DMRT (Duncan’s multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash (FA).

8.1 § 0.054b (24.23)
19.24 § 0.218b (32.14)
22.1 § 0.331b (27.74)

7.66 § 0.0.050c (17.48)
17.62 § 0.288c (21.01)
19.08 § 0.106c (10.28)

(A) Root length (cm)
Pre-flowering
6.52 § 0.048d
Flowering
14.56 § 0.087d
Post-flowering
17.3 § 0.122d

T2
10%FA

0%FA

T1
5%FA

Parameters/stages

T0 control

Table 3. Effect of fly ash on root length (cm/plant), root fresh weight (g/plant) and dry weight (g/plant) at various growth stages of Carthamus tinctorius L.

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Israel Journal of Plant Sciences

30

25

10

100

20

d

5

40
Pre-flowering

d

d

15
d
c

T0

120
d

40
d
c

T0
T1

Flowering
a

35
c
b

T1

Pre-flowering

c
c

b

20
b

T2

140

b

T2
T3

Post-flowering

a
A

c
e
e

a
f

T3

Flowering

80
e

f

5

e
f

0
Treatment

T4

e

T4

T5

a a

Post-flowering

b
B

b
e
f
f

60
a

0

f

Treatment

T5

Figure 1. Effect of fly ash on the number of branches per plant
(A) and the number of leaves per plant (B) at various growth
stages of Carthamus tinctorius L. Values represent mean § SE
(n D 5). Means marked by different letters are significantly (p <
0.05) different from their respective controls along a growth
stage according to DMRT (Duncan’s multiple range test). T0: 0,
T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash (FA).

increase was found in T3 fly ash amendment. However, a
dose-dependent reduction was observed with higher doses
in T4 and T5 and the least reduction was found with 75%
pre-flowering stage. The maximum variation (77.77%)
was recorded with 25% fly ash treatment at pre-flowering
stage followed by flowering (50.48%) and post-flowering
(42%) stages.
The total number of leaves significantly (p < 0.05)
increased by the fly ash amendment at all stages of plant
development (Figure 1B). The highest increase (94.79%)
was observed in T3. However, a dose-dependent reduction
was observed with higher doses (T4, T5). The highest
increase in number of leaves (94.79%) was recorded in T3
with 25% fly ash level at the pre-flowering stage followed
by the flowering (55.66%) and post-flowering (35.25%)
stages.
Fresh and dry biomass of leaves increased in T2 and
T3 at all stages of plant development (Table 4A,B). The
maximum increase in fresh weight of leaves was recorded

Values in parentheses indicate percent variation with reference to respective controls. Mean § SE (n D 5). Values with different superscripts are significantly (p < 0.05) different from each other DMRT
(Duncan’s multiple range test). ). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash (FA).

0.198 § 0.001f (44.84)
1.240 § 0.003f (25.34)
2.972 § 0.0004f (24.85)
0.292 § 0.001e (18.66)
1.434 § 0.003e (13.66)
3.217 § 0.002e (18.65)
0.788 § 0.001a (119.4)
4.780 § 0.003a (187.77)
7.528 § 0.001a (90.34)
0.478 § 0.001c (33.14)
2.551 § 0.007c (53.58)
4.725 § 0.003c (19.46)
(B) Leaf dry weight (g)
Pre-flowering
0.359 § 0.001d
Flowering
1.661 § 0.002d
Post-flowering
3.955 § 0.001d

0.561 § 0.001b (56.26)
3.112 § 0.002b (87.35)
5.747 § 0.009b (45.30)

1.457 § 0.016f (19.27)
4.276 § 0.020f (24.83)
5.731 § 0.051f (27.41)
1.600 § 0.001e (11.35)
4.787 § 0.001e (15.85)
6.137 § 0.0018e (22.27)
3.835 § 0.011a (112.46)
10.349 § 0.001a (81.92)
12.675 § 0.006a (60)
2.082 § 0.04c (13.30)
6.887 § 0.00c (21)
8.828 § 0.00c (11.80)
(A) Leaf fresh weight (g)
Pre-flowering
1.805 § 0.01d
Flowering
5.689 § 0.00d
Post-flowering
7.896 § 0.01d

2.789 § 0.00b (54.51)
7.331 § 0.33b (28.86)
10.964 § 0.07b (38.85)

75%FA
25%FA
Parameters/stages

0%FA

5%FA

10%FA

T4
T3
T2
T1
T0 control

Table 4. Effect of fly ash on the leaf fresh (g/plant) and dry weight (g/plant) at various growth stages of Carthamus tinctorius L.

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50%FA

Qurratul et al.

T5

6

in T3, but a slight increase was seen in T1 at all stages of
plant development. The fresh and dry weight of leaves
was decreased in T4, T5, and the highest decrease occurred
at 75% fly ash application. The maximum increase in
fresh weight of leaves (112.46%) was recorded in T3 at
pre-flowering stage followed by flowering (81.92%) and
post-flowering (60%) stages. A similar trend was found
for leaf dry weight.
Yield parameters
Effects of fly ash treatment on the production of reproductive traits are shown in Figure 2A
D. The number of
heads per plant and seeds per head increased from T1 to
T3. The highest increase was observed in T3. However, a
dose-dependent reduction was observed with higher application rates (T4, T5), and the least reduction was found at
75% fly ash level. The highest increase in the number of
heads per plant (68.91%) was found with 25% fly ash
application followed by 10% (32.43%). The highest
increase (59.70%) in the number of seeds per head
occurred under 25% fly ash application followed by
38.05% at 10% application. Weight of 100 seeds per
plant, and weight of seeds per head increased significantly
(p < 0.05) from T1 to T3 at the harvest stage and the highest increase was observed with 25% fly ash treatment.
However, it declined with higher fly ash dose reaching a
minimum at 75% fly ash amendment. The highest increase
in weight of 100 seeds per plant (19.31%) was recorded
under 25% fly ash application followed by 10% fly ash
treatment (12.59%). The maximum increase in case of
weight of seeds per head (114.28%) was recorded in T3
followed by the 10% fly ash treatment (58.24%).
Biochemical parameters
Photosynthetic pigments. Effects of fly ash application
on the photosynthetic pigments are shown in
Table 5A
D. Chlorophyll a, b, and carotenoid contents
increased under low concentration of fly ash, but
decreased under high application rates (Table 5A
D).
The highest increase in pigment contents was observed
under 25% fly ash level at all stages of plant growth. The
highest increase in chlorophyll a (52.52%) was recorded
with 25% fly ash level at the flowering stage followed by
post-flowering (49.17%) and pre-flowering (30.36%)
stages, respectively. For chlorophyll b, the highest
increase (144.53%) was recorded with T3 at post-flowering, and the largest change (62.62%) of total chlorophyll
content was induced by 25% fly ash addition at postflowering followed by those at flowering and pre-flowering stages. The highest increase of carotenoids (58.45%)
occurred in T3 at post-flowering stage, followed by preflowering (57.81%) and flowering (43.71%) stages
(Table 5D).

30

a

Number of heads/ plant

25
20

A

b
c
d

15

e
f

10
5

Weight of 100 seeds/ plant (g)

Israel Journal of Plant Sciences

0

50
45
40
35
30
25
20
15
10
5
0

T1

T2
T3
Treatment

T4

c
e

T1

T2
T3
Treatment

c
d

e
f

T1

T2

2.5

B

b

T0

C

b

T3

T4

T5

Treatment

a

d

a

T0

T5

T4

f

T5

Weight of seeds/ head (g)

Number of seeds/ head

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T0

5
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0

7

D

a

2

b
c

1.5
d
1

e
f

0.5
0
T0

T1

T2

T3

T4

T5

Treatment

Figure 2. Effect of fly ash on the number of heads per plant (A), number of seeds per head (B), weight of 100 seeds per plant (C) and
weight of seeds per head (D) at harvest of Carthamus tinctorius L. Values represent mean § SE (n D 5). Means marked by different letters are significantly (p < 0.05) different according to DMRT (Duncan’s multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5:
75% fly ash (FA).

Table 5(A). Effect of fly ash on the chlorophyll a content in
leaves (mg/g fresh weight) at various growth stages of Carthamus tinctorius L.

Table 5(B). Effect of fly ash on the chlorophyll b content in
leaves (mg/ g fresh weight) at various growth stages of Carthamus tinctorius L.

Treatment

Treatment

Pre-flowering

T0 control
0%FA
T1
5%FA
T2
10%FA
T3
25%FA
T4
50%FA
T5
75%FA

T0 control
0%FA
T1
5%FA
T2
10%FA
T3
25%FA
T4
50%FA
T5
75%FA

Pre-flowering

Flowering

Post-flowering

1.37 § 0.02d

1.447 § 0.001d

0.905 § 0.002d

1.453 § 0.085c
(6.058)
1.706 § 0.145b
(24.52)
1.786 § 0.121a
(30.36)
1.281 § 0.015e
(6.49)
1.18 § 0.044f
(13.86)

1.510 § 0.001c
(4.35)
1.993 § 0.001b
(37.73)
2.207 § 0.009a
(52.52)
1.437 § 0.001d
(0.69)
1.349 § 0.001e
(6.77)

1.045 § 0.002c
(15.46)
1.208 § 0.001b
(33.48)
1.350 § 0.002a
(49.17)
0.823 § 0.001e
(9.06)
0.727 § 0.026f
(19.66)

Values in parentheses indicate percent variation with reference to respective controls. Mean § SE (n D 5). Values with different superscripts are
significantly (p < 0.05) different from each other by DMRT (Duncan’s
multiple range test). ). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly
ash (FA).

Flowering

Post-flowering

0.252 § 0.055

0.620 § 0.001

d

0.238 § 0.002d

0.321 § 0.001bc
(27.38)
0.350 § 0.001b
(38.88)
0.445 § 0.001a
(76.58)
0.279 § 0.001cd
(10.71)
0.235 § 0.001d
(6.74)

0.668 § 0.002c
(7.74)
0.808 § 0.002b
(30.32)
0.878 § 0.002a
(41.61)
0.456 § 0.005e
(26.45)
0.380 § 0.001f
(38.70)

0.298 § 0.002c
(25.21)
0.439 § 0.002b
(84.45)
0.582 § 0.002a
(144.53)
0.195 § 0.002e
(18.06)
0.156 § 0.001f
(34.45)

cd

Values in parentheses indicate percent variation with reference to respective controls. Mean § SE (n D 5). Values with different superscripts are
significantly (p < 0.05) different from each other by DMRT (Duncan’s
multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly
ash (FA).

Qurratul et al.

Table 5(C). Effect of fly ash on the total chlorophyll content in
leaves (mg/g fresh weight) at various growth stages of Carthamus tinctorius L.

activity in a dose-dependent manner at pre-flowering,
flowering and post-flowering stages (Figure 3B).

Treatment

Pre-flowering

Flowering

T0 control
0%FA
T1
5%FA
T2
10%FA
T3
25%FA
T4
50%FA
T5
75%FA

1.708 § 0.001

2.074 § 0.02

1.164 § 0.017

Soluble sugar content. Low dosages of fly ash amendment increased sugar content of Carthamus tinctorius L.
(Figure 3C). The highest increase was observed with 25%

2.192 § 0.002c
(5.68)
2.814 § 0.002b
(35.67)
3.091 § 0.003a
(49.03)
1.905 § 0.002e
(8.14)
1.73 § 0.003f
(16.58)

1.337 § 0.002c
(14.86)
1.647 § 0.001b
(41.49)
1.893 § 0.027a
(62.62)
1.009 § 0.001e
(13.31)
0.905 § 0.002f
(22.25)

d

Values in parentheses indicate percent variation with reference to respective controls. Mean § SE (n D 5). Values with different superscripts are
significantly (p < 0.05) different from each other by DMRT (Duncan’s
multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly
ash (FA).

Protein content. The soluble protein content in the
leaves increased under low (25%) fly ash application as
compared to control at all the stages of plant growth. The
highest increase was found at the flowering stage
(Figure 3A). Further increase in fly ash concentration
decreased protein content in a dose-dependent manner.
Nitrate reductase activity. NR activity increased in T2
and T3 compared to the non-treated control at all stages of
plant development, but the highest increase was found in
T3 (25%) at the flowering stage (Figure 3B). Higher levels
(50% and 75%) of fly ash application decreased NR
Table 5(D). Effect of fly ash on the carotenoid content in leaves
(mg/g fresh weight) at various growth stages of Carthamus tinctorius L.
Treatment

Pre-flowering

Flowering

T0 control
0%FA
T1
5%FA
T2
10%FA
T3
25%FA
T4
50%FA
T5
75%FA

0.768 § 0.002

0.819 § 0.001

d

0.852 § 0.001
(10.93)
1.009 § 0.001b
(31.38)
1.212 § 0.001a
(57.81)
0.738 § 0.001e
(3.90)
0.656 § 0.004f
(14.58)
c

Post-flowering
d

0.855 § 0.002
(4.39)
1.013 § 0.019b
(23.68)
1.177 § 0001a
(43.71)
0.738 § 0.001e
(9.89)
0.713 § 0.001f
(12.94)
c

0.467 § 0.001

d

0.518 § 0.002
(10.92)
0.637 § 0.002b
(36.40)
0.740 § 0.002a
(58.45)
0.440 § 0.001e
(5.78)
0.418 § 0.002f
(10.49)
c

Values in parentheses indicate percent variation with reference to respective controls. Mean § SE (n = 5). Values with different superscripts are
significantly (p < 0.05) different from each other by DMRT (Duncan’s
multiple range test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly
ash (FA).

Pre-flowering

Flowering

40
Protein content (mg g-1 FW)

1.771 § 0.001c
(3.68)
2.184 § 0.002b
(27.86)
2.468 § 0.002a
(44.49)
1.535 § 0.003e
(10.12)
1.320 § 0.001f
(22.71)

d

35
30
25

b

c

d
d

e
a

b

20

f
e

c

d

15

A

a

b

c

Post-flowering

a

e

f
f

10
5
0
T0

T1

T2

T3

T4

T5

Treatment

Pre-flowering
NR activity (µmol g FW-1h-1)

d

Post-flowering

4.5
4
3.5
3
2.5
2
1.5
1
0.5
0

Flowering
a

Post-flowering

B

b

c

a

d
c

d

e

b

f

e
d

c

b

T0

T1

T2

f

a

T3

e

f

T4

T5

Treatment

Pre-flowering
Sugar content (mg gFW-1)

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8

100
90
80
70
60
50
40
30
20
10
0

Flowering

Post-flowering

C

a

b
c
d

a

b
c
d
d

T0

c

T1

b

e

a

f

e
e

T2
T3
Treatment

T4

f

f

T5

Figure 3. Effect of fly ash on soluble protein content (A), nitrate
reductase (NR) activity (B) and soluble sugar content (C) at various growth stages of Carthamus tinctorius L. Data are means §
SE (n D 5). Values marked by different letters are significantly
(p < 0.05) different from their respective controls along a
growth stage according to DMRT (Duncan’s multiple range
test). T0: 0, T1: 5, T2:10, T3:25, T4: 50 and T5: 75% fly ash
(FA).

Israel Journal of Plant Sciences
fly ash application at the flowering stage. Under high
application rates, a dose-dependent decrease in sugar content was found (Figure 3C).
Discussion

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Physicochemical properties of the soil
The physicochemical properties of fly ash depend on a
number of factors such as the nature of the parent coal,
conditions of combustion, type of emission control devices and storage and handling methods (Ahmaruzzaman
2010). The physiochemical analysis of 75% fly ash
amended soil used in the present study demonstrates a rise
in alkalinity (pH 7.38 to 7.86), EC 1.23, WHC 59.54 and
lowered BD from 1.58 to 1.35 (Table 1). Singh et al.
(2012) also found similar increases in pH, EC, and WHC
and reduced BD in fly ash amended soil. According to
Khan and Khan (1996), the increase in soil pH might be
due to the neutralization of H+ by alkali salts as well as to
solubilization of basic metallic oxides of fly ash in the
soil. Gupta et al. (2007) depicted that the increased soil
pH might be a result of precipitation of soluble cations in
the fly ash soil. Increase of EC values due to fly ash could
suggest that the binding of metal ions to soil particles
occurred readily, increasing the availability of metal
nutrients to the growing plants (Mishra et al. 2007). The
increase in WHC was due to greater space between the
soil particles (Khan & Khan 1996). Sharma and Kalra
(2006) found a general decrease in soil bulk density following fly ash addition. This in turn improved soil porosity and enhanced water retention capacity. Organic carbon
was also increased with fly ash amendment. Gradual
increase in pH, EC, WHC, OC, available P, K, and S, and
a decrease in BD and total N content of the soil were
observed (Table 1). Similar findings were also reported in
previous studies by different workers (Singh & Siddiqui
2003; Gond et al. 2013).
Morphological parameters
Plant growth analysis is a necessary step towards understanding plant performance and productivity (Parween
et al. 2011a). The present study demonstrated that lower
concentration (25%) of fly ash is effective, non-phytotoxic
and seemed to elevate the growth parameters up to a limit.
At higher amendment levels, shoot length, fresh and dry
weight of shoot (Table 2A
C), root length, fresh and dry
weight of the root (Table 3A
C), number of branches and
leaves (Figure 1A,B), fresh and dry weight of leaves
(Table 4A,B) were reduced in all growth stages evaluated.
Our results are in accordance with other studies which
demonstrated growth stimulation under low application
levels (Katiyar et al. 2012; Singh et al. 2012). The fly ash
used in the present study contains high concentration of
K. When added to soil in moderate amounts (25%), it

9

improved plant performance, but had deleterious effects
under higher application rates. No visible symptoms of
nutrient deficiency or phytotoxicity were observed at this
study.
Wong and Wong (1990), with Brassica chinensis and
B. parachinensis, and Pandey et al. (1994), with Helianthus annuus, also reported beneficial effects of low concentrations of fly ash. Recently, Gupta and Sinha (2006)
studied the effect on different amendments of fly ash on
the growth of Brassica juncea. They reported improved
shoot but not root growth by amendment of 25% fly ash.
Stimulation of plant growth at lower soil applications of
fly ash suggests that certain essential elements, such as
Ca, Mg, K, Mn, Mo, Zn, or S present in the fly ash, were
made more readily available to the plants (Singh et al.
1997). At higher application rates, the resultant high soil
pH might have reduced the extractability and availability
of some nutritional mineral elements, thus reducing plant
performance (Wong & Wong 1989).
The highest reduction of shoot length (36.12%) was
recorded in T5 (75%) at the pre-flowering stage, followed
by the post-flowering (10.86%), and the flowering
(7.63%) stages (Table 2A). However, root length
decreased with the increase in concentration of fly ash
(50% and 75%) (Table 3A). Inhibition of root growth may
be due to compaction of fly ash particles which probably
served as physical barrier to root elongation. Fly ash contains a high level of heavy metals, which may inhibit root
growth via an effect on cell division rate (Singh et al.
2008). Roots are the first organs that come in contact with
the toxic metals in soil and most of the toxic metals can
be deposited in the root tissues. Metal deposition in the
root may restrict movement of the toxic metals to the
leaves and other shoot organs (Gupta et al. 2002), thereby
regulating to some extent the response of the shoot to the
fly ash constituents. Our results demonstrated growth
reduction under higher concentrations of fly ash. Another
reason for this decrease may have been induced changes
in the chemical properties of the soil (Mishra & Shukla
1986; Wong & Wong 1989). Previous experiments, both
under field and greenhouse conditions, with fly ash
amendments showed that different chemical constituents
of fly ash can promote plant growth and improve the agronomic properties of the soils (Singh et al. 2012). Singh
et al. (1997) reported that 5
10% fly ash amendments to
the soil enhanced seed germination and seedling growth,
but higher applications (20% and 30%) either delayed or
drastically inhibited plant growth and development.
Dwivedi et al. (2007) found optimum growth response in
different rice cultivars with 25% fly ash amendment. The
present study demonstrates that the lower plant biomass at
higher level of fly ash might be due to accumulation of
heavy metals. This is in agreement with an earlier report
on leguminous plants (Vajpayee et al. 2000). Sharma
et al. (2008) have also reported the reduction in root,

10

Qurratul et al.

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stem, leaf and total plant biomass of Daucus carota L.
with combined treatments of Cd and Zn due to accumulation of Cd and Zn. Similarly, in the present study stimulated accumulation could have induced growth cessation
in Carthamus tinctorius L.
The reduced number of leaves with the increase in fly
ash concentration may also be due to reduced chlorophyll
content and inhibited photosynthates synthesis. The
results of Parween et al. (2011a) suggest the same for
Vigna radiata L.
Yield parameters
Yield is the most important parameter from commercial
point of view and numerous studies have shown a yield
increase under fly ash amendment (Gupta et al. 2002; Jala
& Goyal 2006). In our study, 25% fly ash increased yield
parameters, i.e. number of heads/plant, number of seed/
head, weight of 100 seed/plant and weight of seed/head
(Figure 2A
D). However, as in other crops, the observed
decline in yield at higher fly ash levels may be linked to
apparent metal toxicity caused by accumulation of Mn,
Ni, Co, Zn, Cu, Pb, Cr and Cd in plant tissues (Mishra
et al. 2007). Lee et al. (2006) reported that an addition of
around 90 mg/ha fly ash in paddy field soil can significantly increase the grain yield in Korean rice cultivars.
Dwivedi et al. (2007) found that the grain weight in three
Indian rice cultivars was highest with 25% fly ash amendments. Singh and Agrawal (2010) also reported in
increase in economical yield in mung bean plants with
5
10% fly ash amendments. Pandey et al. (2010) found
that the chickpea yield decreased significantly at 50% and
100% fly ash dose; however, at 25% fly ash treatment the
yield was not affected. Su and Wong (2002) found that
the addition of an ash
sludge mixture significantly
improved the dry weight yields of corn seedling as compared to the control. They also emphasized that the lower
amendments of ash
sludge mixture (5% and 10%) at
1:5 v/v resulted in the highest dry weight yields. Our
results are consistent with the result of Katiyar et al.
(2012), demonstrating a positive impact of a lower concentration fly ash on various morphological and yield
parameters at different developmental stages, and a
respective decline under higher concentration of fly ash.
Because fly ash lacks nitrogen, its application at higher
concentrations results in severe deficiency of nitrogen in
the soil and the plant resulting in suppressed growth and
yield (Gupta et al. 2002).
Biochemical parameters
Photosynthesis is the ultimate physiological limitation to
crop production. The photosynthetic capacity of individual leaves is a prime factor that determines crop dry matter and yield. Photosynthetic efficiency depends on leaf

area, chlorophyll contents and stomatal response to the
environment (Ayari et al. 2000). Generally, plant tolerance to fly ash is reflected in chlorophyll and carotenoid
contents. Chlorophyll and carotenoid contents are indicators of environmental stress-induced damage to the photosynthetic system (Shoresh et al. 2011; Pandey 2013). In
the present study, fly ash at high concentrations (50%,
75%) induced a decrease in total pigment concentration
(Table 5A
D). Chlorophyll suppression may result from
restriction of the level of precursors or through targeting
-SH groups of aminolevulinic acid dehydratase in the
presence of metals (Gupta et al. 2000). Accumulation of
heavy metals leads to inhibition of chlorophyll formation
(Krupa and Baszynski, 1995), resulting in a decline in
chlorophyll content. The breakdown of photosynthetic
pigment may be due to substitution of Mg2+ ions in chlorophyll molecules by metal ions such as Cu2+, Zn2+, Cd2+,
Pb2+, Ni2+ (Kupper et al. 1998).
The carotenoids are naturally occurring non-enzymatic antioxidants in plants that serve as accessory pigments for photosynthesis. They protect the chlorophyll
molecule under environmental stress conditions by different mechanisms such as quenching the photodynamic
reactions, and avoiding peroxidation and thereby membrane damage (Kenneth et al. 2000). Our results show that
the carotenoid content was decreased with increasing concentration of fly ash and an enhanced level of carotenoid
with 25% fly ash application might have occurred to protect the plant against the toxic effect of free radicals. In
support of the present results, an increase in carotenoids
was also reported in Sesbania cannabina and chickpea
grown on different fly ash doses (Sinha & Gupta 2005;
Pandey et al. 2010) and Fenugreek grown on different
amendment of tannery sludge (Sinha et al. 2007), which is
considered a defense strategy of plants to combat metal
stress. These findings are also in agreement with results
reported by Robab et al. (2010) and Gond et al. (2013) for
photosynthetic pigments in Solanum nigrum L. and Solanum melongena, respectively.
In our study, protein content significantly increased
with lower (25%) fly ash application. However, reduction
in protein content (Figure 3A) was also observed in this
study under higher levels of fly ash. Decline in protein
may result from an increase in the activity of protease or
other catabolic enzymes. Similar results were reported
earlier by Sharma et al. (2010), with Pisum sativum L.
and Gautam et al. (2012), with Brassica juncea. RomeroPuertas et al. (2002) also reported that fly ash and tannery
sludge having large amounts of heavy metals induce production of reactive oxygen species in plants. These free
radicals damage the photosynthetic apparatus and degrade
proteins through oxidative modification and increase in
proteolytic activity.
Nitrate reductase (NR) is an induced enzyme that regulates the first step of nitrogen metabolism (Abrol et al.

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Israel Journal of Plant Sciences
1976). In the present investigation, the NR activity
(Figure 3B) decreased with higher concentrations of fly
ash, possibly due to decomposition of the enzyme molecule. Because fly ash is deficient in nitrogen, inhibition of
NR activity may have been caused either by the reduction
in the content of the protein, or by affecting its substrate
availability. Inhibition of NR activity was also reported in
Leuacaena leucocephala at a higher level of fly ash by
Gupta et al. (2000). NR inhibition may also be due to the
binding ability of these metals to -SH groups of enzymes
(Gupta et al. 2000).
Sugars are essential for normal NR function (Campbell 1999). In the present study, leaf sugar content
increased by the 25% of fly ash treatment (Figure 3C) and
decreased with further increase in fly ash application. The
decline in sugar content correlated with the reduction of
pigment concentrations. This suggests that the photosynthesis machinery could have been damaged and thus supports the possibility of an indirect effect of fly ash
application, i.e. damage to the photosynthesis machinery
and sugar supply and consequently affecting NR activity.
Activity of nitrate reductase is directly related to nitrate
concentration which in turn plays an important role in protein synthesis. In the present study, maximum biochemical activities were observed at the flowering stage and
later declined sharply. This phenomenon occurs mainly
because aging leaves have lower metabolic activities compared to younger leaves, as aging is a period of catabolism, which may cause decreased biochemical activity
(Haddad et al. 2009; Parween et al. 2011b).

Conclusion
The present study demonstrates stimulating effects of fly
ash, on growth, biomass production, productivity and photosynthetic pigments on safflower growing on soil
amended with low (25%) levels of fly ash. Higher percentages reduced plant performance, suggesting detrimental
effects of fly ash.

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