International Journal of Agricultural

Science and Research (IJASR)

ISSN(P): 2250-0057; ISSN(E): 2321-0087
Vol. 6, Issue 5, Oct 2016, 197-206
TJPRC Pvt. Ltd

EFFECT OF DIFFERENT COMBINATIONS OF CHEMICAL FERTILIZERS

(UREA, SSP, MOP, NP, NK, NPK) AND BIOFERTILIZER ON MDA, PEROXIDASE
ACTIVITY AND PROLINE CONTENT IN WHEAT (TRITICUM AESTIVUM L.)
SEEDLINGS
SANDHYA SHARMA & ANGURBALA BAFNA
Department of Biochemistry, Government Holkar Science College, Indore, Madhya Pradesh, India
ABSTRACT
Normal growth and development of plants is greatly dependent on the capacity to overcome abiotic and biotic
stresses. These stresses prevent plants from realizing their full genetic potential and limits food production. Fertilizers
are sources of plant nutrient that can be added to soil to maintain its natural fertility. The application of chemical
fertilizers over a period resulted in poor soil health, reduction in produce and increase in incidences of pest and disease
and environmental pollution. Bio-fertilizers are eco-friendly, effective and economical alternate of chemical fertilizers.
An antioxidant enzyme Peroxidase has important role in the metabolic reactive oxygen species (ROS) and defence
The objective of this study was to evaluate the effects of chemical fertilizers (N, P, K) in six different combination
(N, P, K, N+P, N+K and N+P+K) and combinations with biofertilizers N+A, P+A, K+A, NP+A, NK+A, NPK+A on the
peroxidase enzyme and stress markers in 7th day old wheat seedling. Results showed that the activity of peroxidase
enzyme, MDA and proline content was significantly different (p<0.05) between control and fertilizer treatments.
The lowest value of MDA with NK and NPA combinations, proline with NK and KA combinations and peroxidase activity
with NPK and NKA combinations were observed. Low level of MDA, proline and peroxidase indicates that wheat

Original Article

against oxidative stress damage. MDA and proline are the stress markers increase in plant cells as a response to stresses.

seedlings do not face any type of stress in presence of chemical fertilizers as well as biofertilizer.
KEYWORDS: Fertilizers, Urea, SSP, MOP, Triticum Aestivum L., Azotobacter, Peroxidase, Proline, MDA.

Received: Aug 19, 2016; Accepted: Sep 15, 2016; Published: Sep 20, 2016; Paper Id.: IJASROCT201623

INTRODUCTION
For optimum plant growth, nutrients must be available in sufficient and balanced quantities. Soil contains
natural reserves of plant nutrients, but these reserves are largely in forms unavailable to plants, and only a minor
portion is released each year through biological activity or chemical processes. This release is too slow to
compensate for the removal of nutrients by agricultural production and to meet crop requirements (Chen J., 2006).
Fertilizers are sources of plant nutrient that can be added to soil to maintain its natural fertility. The chemical
fertilizer increased the production but also produce many harmful effects. The continued use of chemical fertilizers
causes health and environmental hazards such as ground and surface water pollution by nitrate leaching
(Pimentel 1996). Therefore the use of biofertilizer in place of chemical fertilizer is both economical and
environmental friendly. The use of living bacteria (biofertilizer) accelerates mineralization of organic residues in
soil, therefore makes the nutrients more available. At the same time due to effect of living bacteria from
biofertilizer, the uptake of heavy metals decreased, indicated that use of bio-fertilizers instead of chemical

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Sandhya Sharma & Angurbala Bafna

fertilizers is not sufficient whereas using of bio-fertilizers increased the efficiency of chemical fertilizers
(Lvai L. et al., 2008). Normal growth and development of plants is greatly dependent on the capacity to overcome abiotic
and biotic stresses. A common denominator in stress conditions is the production of reactive oxygen species (ROS) within
different cellular compartments of the plant cell (Jithesh M. N. et al., 2006). Excessive production of ROS results in
progressive oxidative damage and ultimate cell death. Despite their destructive activity, they are well-described second
messengers in a variety of cellular processes, including conferment of tolerance to various environmental stresses. Whether
ROS would serve as signalling molecules or could cause oxidative damage to the tissues depends on the delicate
equilibrium between ROS production, and their scavenging. Efficient scavenging of ROS produced during various
environmental stresses requires the action of several non-enzymatic as well as enzymatic antioxidants present in the tissues
(Sharma P. et al., 2012). Therefore, the objective of present study is to evaluate the effects of different combinations of
chemical and bio-fertilizers on peroxidase activity, MDA and proline content in wheat (Triticum aestivum L. GW 366)
seedlings.

MATERIALS AND METHODS

Soil: The basic requirement for this experiment was soil. Soil was collected from Agriculture college campus
land. Stones and other hard material removed from it. It was then grinded and filtered. This finely powdered soil was then
filled with uniform layer in the Petri plates of equal size. The soil in the Petri plates was moistened with water. Sowing of
seeds was done in this soil after seed treatment.
Fertilizers Solution: For study, recommended dosage of some selected chemical fertilizers are Urea, SSP and
MOP were used.
Biofertilizers: Azotobacter was used.
Seeds: Seeds of wheat of uniform size were selected and surface sterilized with 0.1% solution of mercuric
chloride for 5min to avoid any fungal growth, followed by washing for 4-5 times with distilled water.
Preparation of Soil: After collection of soil, 75g soil were weighed and put into Petri-dish and then add the
recommended dose of chemical fertilizers (120:60:30 Kg/ha) in following different combinations and add 45 ml of distilled
water in it. Keep the Petri dishes for 7 days in dark to generate field capacity. The experiment was completely randomized
and consisted of 13 treatments in triplicate. Combinations were control (without fertilizers), Urea (N), SSP (P), MOP (K),
Urea + SSP (N+P), Urea + MOP (N+K), Urea + SSP + MOP (N+P+K), N+A, P+A, K+A, NP+A, NK+A, NPK+A.
For biofertilizer slurry of Azotobacter (5-10 g/kg of seeds) was prepared with distilled water and the overnight
soaked seeds mixed in this slurry and dried in shade. Then 12 seeds were sowed in each type of combination.
All the Petri plates were kept at room temperature (255oC) for 7 days. During this period all plates were kept in
sun light for 6 hours. On the 7th day peroxidase activity, MDA and proline content were measured under standard
condition.
Malondialdehyde Estimation: It is estimated by following the method of Heath R. L. and Packer L. (1968).
Method

The tissue was homogenized with 5% TCA and 1 ml of homogenate was mixed with 4 ml of TBA reagent.

Impact Factor (JCC): 4.8136

NAAS Rating: 3.53

Effect of Different Combinations of Chemical Fertilizers (Urea, SSP, MOP, NP, NK, NPK) and
Biofertilizer on MDA, Peroxidase Activity and Proline Content in Wheat (Triticum Aestivum L.) Seedlings

199

The reaction mixture were heated at 950C for 30 minutes in a water bath and then quickly cooled in an ice bath
and centrifuged at 1900 g for 10 min.

The absorbance of the colored supernatant was measured at 532 nm.

For the reference blank 1 ml of 5% TCA was mixed with 4 ml TBA reagent.

Calculation
Concentration of MDA was calculated by using the extinction coefficient of 155 mM-1cm-1 for MDA at 532 nm
using formula
A= lc
Peroxidase Activity: Peroxidase activity was measured according to Summer J. B. et al., (1943).
Extraction:

Homogenize the material in ice cold 0.1 M phosphate buffer, pH 6 in a chilled pestle and mortar.

Strain thorough the two muslin cloth and centrifuge the homogenate at 16000g for 20 min. at 40C. Use the
supernatant as enzyme source.

Enzyme Assay

Pipette out 1 ml of o-dianisidine, 0.5 ml of H2O2, 1 ml of phosphate buffer and 2.4 ml distilled water into test
tube.

For blank, exclude H2O2 but add additional volume of water.

Incubate at 300C and start the reaction by adding 0.2 ml of enzyme.

After 5 min, stop the reaction by adding 1 ml of 2N H2SO4.

Read the absorbance at 430 nm.

Calculation
Express the specific activity of enzyme as unit/min/mg of protein or per g of weight of sample considering one
unit of enzyme as increase in OD by 1.0 under standard condition.
Proline Estimation: Proline was measured according to Bates L. S. et al., (1973).
Extraction:

Extract 0.5g of plant material by homogenizing in 10 ml of 3 % aqueous sulphosalycylic acid.

Heat it in the boiling water bath for 1 h.

Terminate the reaction by placing the tube in ice bath.

Add 4 ml toluene to the reaction mixture and stir well for 20-30 sec.

Separate the chromophores containing toluene layer with the help of separating funnel and warm to room
temperature.

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200

Sandhya Sharma & Angurbala Bafna

Measure the red colour intensity at 520 nm in spectrophotometrically against toluene blank.

Run a series of standard with pure proline in a similar way and prepare a standard curve.

Find out the amount of proline in the test sample from the standard curve prepared from pure proline and express
on fresh weight basis of sample.

Calculation
Express the proline content on fresh weight basis as follows:
moles per g tissue = g proline /ml ml toluene / 115.5 5/g of sample

RESULTS AND DISCUSSIONS

MDA Estimation
Table 1 (a): Showing the Effect of Different Combination of Urea with
SSP and MOP on MDA ( Moles/g) in Triticum Aestivum L.
Mean
P Value
P Value
P Value
P Value
MDA Content
SD
W.R.T.
W.R.T.
W.R.T.
W.R.T
( moles/g)
Control
Urea
SSP
MOP
1
Control
11.93
0.20
2
Urea
4.63
0.75
0.000
3
SSP
2.73
0.66
0.000
0.02*
4
MOP
9.56
1.87
0.05*
0.007**
0.002**
5
Urea+SSP
3.93
1.28 0.0002***
0.2 ns
0.1 ns
6
Urea+MOP
2.13
0.55
0.000
0.005**
0.001**
7
Urea+SSP+MOP
7.76
0.40
0.000
0.002** 0.0002***
0.09 ns
NOTE: * for significant, ** for highly significant, *** extremely significant, ns for insignificant.
S. N.

Combinations of
Fertilizers

As shown in table 1 (a) the MDA level in untreated seedlings of wheat was 11.930.20 while in seedlings treated
with Urea, SSP, MOP, N+P, N+K, NPK were 4.630.75, 2.730.66, 9.561.87, 3.931.28, 2.130.55 and 7.760.40
respectively. Decrease in MDA was observed in all combinations as compared to control. These decrease were extremely
significant (p<0.001) in Urea, SSP, NP, NK and NPK and significant (p<0.05) in MOP as compared to control. As
compared to Urea decrease in MDA was highly significant (p<0.01) in MOP, NK and NPK combinations and significant in
SSP and insignificant in NP combination. MDA was extremely significantly increased in NPK combination and highly
significantly increase in MOP while Insignificant in NP combination as compared to SSP. As compared to MOP decrease
in MDA was highly significant in NK and insignificant in NPK combination.
In the present study highest decrease in MDA level was observed in NK combination as compared to control. As
compared to Urea highest change in MDA was observed in NPK combination. Highest change was observed in NPK as
compare to SSP and in NK as compared MOP.

Impact Factor (JCC): 4.8136

NAAS Rating: 3.53

Effect of Different Combinations of Chemical Fertilizers (Urea, SSP, MOP, NP, NK, NPK) and
Biofertilizer on MDA, Peroxidase Activity and Proline Content in Wheat (Triticum Aestivum L.) Seedlings

201

Table 1 (b): Showing the Effect of different Combinations of Chemical and

Biofertilizers on MDA ( Moles/g) in Triticum Aestivum
S.
N.

Combinations of
Chemical &
Biofertilizer

1
2
3
4
5
6
7

Control
Urea+A
SSP+A
MOP+A
Urea+SSP+A
Urea+MOP+A
Urea+SSP+MOP+A

Mean
MDA
Content
(moles/g)
11.93
11.05
9.53
9.66
8.8
10.5
9.16

SD
0.20
0.51
0.57
0.76
0.4
1.6
0.56

P Value
WRT
Control

P Value

0.3 ns
0.001***
0.03*
0.0001***
0.1 ns
0.0007***

0.0003***(a)
0.000***(b)
0.5 ns(c)
0.03*(d)
0.04*(e)
0.08 ns(f)

P Value
WRT NA

P Value
WRT PA

0.02*
0.08 ns
0.006**
0.4 ns
0.01**

0.4 ns
0.07 ns
0.2

ns

P Value
WRT KA

0.3 ns
0.3 ns

NOTE : * for Significant, ** for Highly Significant, *** Extremely significant, ns for Insignificant, (a) for
N/NA, (b) for SSP/PA, (c) for MOP/KA, (d) for NP/NPA, (e) for NK/NKA, (f) for NPK/NPKA.
As shown in table 1 (b) the MDA level in untreated seedlings of wheat was 11.930.20 while the MDA level of
seedlings treated with NA, PA, KA, NPA, NKA, NPKA were 11.050.51, 9.530.57, 9.660.76, 8.80.4, 10.51.6 and
9.160.56 respectively. Decrease in MDA was observed in all combination as compared to control. This decrease was
extremely significant (p<0.001) in PA, NPA, and NPKA combinations and significant (p<0.05) in KA combination while
insignificant in NA and NKA combinations compared to control. Increase in MDA level was observed extremely
significant in NA and PA combinations as compared to Urea and SSP insignificant in KA and NPKA combinations as
compare to MOP and NPK combinations. As compared to NA significant change was observed in PA and highly significant
in NPA and NPKA combinations and insignificant in KA and NKA. Change was insignificant in KA, NPA and NPKA as
compared to PA and as compare to KA insignificant change was observed in KA and NPKA combinations.
In the present study Triticum Aestivum L. showed highest decrease in MDA with NPA combination as compared
to control. Highest Increase in MDA was observed in PA as compared to SSP. Highest decrease in MDA was observed in
NPA as compare to NA combination. Due to continuous cropping soil may be deficient in essential nutrients which creates
oxidative stress on seedlings but after addition of fertilizers this stress can be relieved that is why in present study low
value of MDA was observed in treated seedlings as compared to control.

Graph 1: Showing the Effect of Chemical and Biofertilizer on MDA content of Triticum Aestivum L.
Peroxidase Activity

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Sandhya Sharma & Angurbala Bafna

Table 2 (a): Showing the Effect of different Combination of Urea with SSP and
MOP on Peroxidase Activity (units/m in/g) in Triticum Aestivum L.
S.N.

Combinations
of Fertilizers

Mean
(in units/min/g)

SD

P Value
W.R.T.
Control

P Value
W.R.T.
Urea

P Value
W.R.T.
SSP

P Value
W.R.T
MOP

1
2
3
4
5
6

Control
42.06
3.91
Urea
39.02
2.12
0.2 ns
SSP
27.98
0.56
0.004**
0.0005***
MOP
36.58
0.20
0.07 ns
0.08 ns
ns
Urea+SSP
36.62
0.53
0.09
0.2 ns
0.000***
ns
Urea+MOP
41.51
1.8
0.4
0.1 ns
0.006**
Urea+SSP+M
7
20.44
0.77
0.0008***
0.000***
0.000***
0.000***
OP
NOTE: * for significant, ** for highly significant, *** extremely significant, ns for insignificant.
As shown in table 2 (a) the peroxidase activity in untreated seedlings of wheat was 42.063.91while in seedlings
treated with Urea, SSP, MOP, N+P, N+K, NPK were 39.022.12, 27.980.56, 36.580.20, 36.620.53, 42.720.53 and
20.440.77 respectively. Decrease in peroxidase activity was observed in all combinations as compared to control. These
decrease were extremely significant (p<0.001) in NPKA combination and highly significant in PA while insignificant in
urea, MOP, NP and NK combinations as compared to control. As compared to Urea decrease in peroxides activity was
observed extremely significant in SSP and NPK combinations and insignificant in MOP, NP and NK combination.
Extremely significant decrease in peroxidise activity was observed in NP and NPK as compared to SSP. As compared to
MOP decrease in peroxidise activity was highly significant in NK and extremely significant in NPK combination.
In the present study highest decrease in peroxidase activity was observed in NPK combination as compared to
control, Urea, SSP and MOP. Decrease in peroxidase activity in NPK combination suggests that NPK reduces the oxidative
stress as compare to control and other fertilizer combinations. Similar work was done by Sabrina B. et al., (2011). They
reported that catalase and ascorbate peroxidase activity was decreased at low level (10 MM) of NPK in roots of wheat
Triticum Durum as compare to control.
Table 2 (b): Showing the Effect of different Combinations of Chemical and
Biofertilizers on Peroxidase Activity (in Units/min/g) in Triticum Aestivum
S. N.

Combinations of
Chemical &
Biofertilizer

1
2
3
4
5
6
7

Control
Urea+A
SSP+A
MOP+A
UREA+SSP+A
UREA+MOP+A
UREA+SSP+MOP+A

Mean
Peroxidase
Activity
(Units/min/g)
42.06
39.16
41.34
37.27
34.6
33.85
38.32

SD
3.91
2.73
0.80
0.37
0.81
0.26
0.36

P Value
WRT
Control

P Value

0.2 ns
0.4 ns
0.08 ns
0.03*
0.02*
0.1 ns

0.5 ns (a)
0.000***(b)
0.08 ns(c)
0.02*(d)
0.002**(e)
0.0009***(f)

P Value
WRT NA

0.1 ns
0.1 ns
0.03*
0.01**
0.3 ns

P Value
WRT PA

P Value
WRT KA

0.0007***
0.0003***
0.002**

0.000
0.01**

NOTE: * for significant, ** for highly significant, *** extremely significant, ns for insignificant, (a) for
N/NA, (b) for SSP/PA, (c) for MOP/KA, (d) for NP/NPA, (e) for NK/NKA, (f) for NPK/NPKA.
As shown in table 2 (b) the peroxidase activity in untreated seedlings of wheat was 42.0633.91 while the
peroxides activity of seedlings treated with Urea+A, SSP+A, MOP+A, N+P+A, N+K+A, NPK+A were 39.162.73,
41.340.80, 37.270.37, 34.60.8, 33.850.26 and 38.320.36 respectively. Decrease in peroxidase activity was observed
in all combination as compared to control. Decrease was significant (p<0.05) in NPA and NKA combinations while
Impact Factor (JCC): 4.8136

NAAS Rating: 3.53

Effect of Different Combinations of Chemical Fertilizers (Urea, SSP, MOP, NP, NK, NPK) and
Biofertilizer on MDA, Peroxidase Activity and Proline Content in Wheat (Triticum Aestivum L.) Seedlings

203

insignificant in NA, PA, KA and NPKA combinations as compared to control. Increase in peroxidase activity was
extremely significant (p<0.001) in PA and NPKA combinations as compared to SSP and NPK combinations respectively.
Decrease was highly significant in NKA as compared to NK combination. Significant decrease was observed in NPA as
compared to NP combination. As compared to NA decrease was highly significant in NKA and significant in NPA
combination while insignificant in PA, KA and NPK combinations. Decrease was extremely significant in KA and NPA
and highly significant in NPKA as compared to PA combination. As compared to KA decrease was extremely significant in
NKA while highly significant increase was observed in NPKA combination.
In the present study Triticum aestivum L. showed highest decrease in peroxidase activity in NKA combination as
compared to control. Highest Increase in peroxidase activity was observed in NPKA as compared to NPK combination.
Highest decrease was observed in NKA as compared to NA combination. Highest decrease in peroxidase activity was
observed in NPA and NKA as compared to PA and KA combinations respectively. Increase in peroxidase activity in NPKA
as compared to NPK combination may be due to the presence of Azotobacter. It is free living nitrogen fixing bacteria
available nitrogen to the plant. This high amount of nitrogen may generate toxicity in plants.

Graph 2: Showing the Effect of Chemical and Biofertilizer on Peroxidase Activity of Triticum Aestivum L.
Table 3 (a): Showing the Effect of Different Combination of urea with
SSP and MOP on Proline (moles/g) in Triticum Aestivum L.
Mean
P Value
P Value P Value P Value
Proline
SD
W.R.T.
W.R.T. W.R.T.
W.R.T
(moles/g)
Control
Urea
SSP
MOP
1
Control
117.05
5.98
2
Urea
107.26
9.11
0.1 ns
3
SSP
87.36
3.9
0.002**
0.01**
4
MOP
96.01
2.6
0.01**
0.08 ns
0.06 ns
5
UREA+SSP
93.85
9.73
0.02*
0.07 ns
0.2 ns
6
UREA+MOP
83.04
4.49 0.001*** 0.007**
0.03*
7
UREA+SSP+MOP
97.74
1.49 0.006**
0.07 ns 0.007**
0.7 ns
NOTE: * for significant, ** for highly significant, *** extremely significant, ns for insignificant.
S.N.

Combinations of
Fertilizers

As shown in table 3 (a) the proline content in untreated seedlings of wheat was 117.055.98 while in seedlings
treated with Urea, SSP, MOP, N+P, N+K, NPK were 107.269.11, 87.373.9, 96.022.6, 93.859.73, 83.044.49 and
97.741.49 respectively. Decrease in proline was observed in all combinations as compared to control. These decreases
were extremely significant in NK combination and highly significant in SSP, MOP and NPK combinations and significant
in NP while insignificant in Urea as compared to control. As compared to Urea decrease was highly significant in SSP and
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Sandhya Sharma & Angurbala Bafna

NK combinations while insignificant in MOP, NP and NPK combinations. Increase in proline was observed highly
significant in NPK combination as compared to SSP. Significant decrease in proline was observed in NK combination as
compared to MOP. In the present study highest decrease in proline was observed in NK combination as compared to
control. Highest decrease was observed in NK combination as compared to Urea and MOP. Significant increase was
observed in NPK combination as compared to SSP.
Table 3 (b): Showing the Effect of Different Combinations of Chemical and
Biofertilizers on Proline (moles/g) in Triticum Aestivum L.
S.N.
1
2
3
4
5
6
7

Combinations of
Chemical and
Biofertilizers
CONTROL
UREA+A
SSP+A
MOP+A
UREA+SSP+A
UREA+MOP+A
UREA+SSP+MOP+A

Mean
Proline
(moles/g)
117.055
91.69
91.69
78.715
90.82
95.15
94.28

SD
5.99
8.1
3.96
5.40
4.49
2.99
5.40

P Value
WRT
Control

P Value

0.006**
0.003**
0.0009***
0.003**
0.004**
0.006**

0.04*(a)
0.1 ns(b)
0.01**(c)
0.3 ns(d)
0.1 ns(e)
0.2 ns(f)

P Value
WRT NA

P Value
WRT PA

0.4 ns
0.05*
0.4 ns
0.2 ns
0.3 ns

P Value
WRT
KA

0.01**
0.4 ns
0.3 ns

0.005**
0.01**

NOTE: * for Significant, ** for Highly Significant, *** Extremely Significant, ns for Insignificant, (a) for
N/NA, (b) for SSP/PA, (c) for MOP/KA, (d) for NP/NPA, (e) for NK/NKA, (f) for NPK/NPKA.
As shown in table 3 (b) the proline in untreated seedlings of wheat was 117.055.98 while the proline of seedlings
treated with Urea+A, SSP+A, MOP+A, N+P+A, N+K+A, NPK+A were 91.6910.8, 91.963.96, 78.725.4, 90.824.49,
95.152.99 and 94.285.40 respectively. Decrease in proline was observed in all combinations as compared to control.
These decrease were extremely significant in KA and highly significant in NA, PA, NPA, NKA and NPKA combinations as
compared to control. Significant change was observed in KA as compared to MOP. Significant decrease was observed in
KA as compared to NA and PA combinations. Highly significant increase was observed in NKA and NPKA combinations
as compared to KA combination.
In the present study highest decrease was observed in KA combination as compared to control. Significant change
was observed in KA as compared to MOP. Significant decrease was observed in KA as compared to NA and PA
combinations. Highest decrease was observed in NKA as compared to KA combination.
It might be due to the control soil was insufficient in nutrient, create stress condition in seedling. Application of
fertilizer might have supply the sufficient amount of nutrient to the soil and these nutrients reduce the stress condition. This
was observed in form of decrease level of proline.

Impact Factor (JCC): 4.8136

NAAS Rating: 3.53

Effect of Different Combinations of Chemical Fertilizers (Urea, SSP, MOP, NP, NK, NPK) and
Biofertilizer on MDA, Peroxidase Activity and Proline Content in Wheat (Triticum Aestivum L.) Seedlings

205

Graph 3: Showing the Effect of Chemical and Biofertilizer on Proline (moles/g) of Triticum Aestivum L

CONCLUSIONS
NPK which stands for nitrogen (promotes leaf growth), phosphorus (root, flower, and fruit), and potassium
(stem and root growth and protein analysis). Nitrogen, Phosphorus and Potassium are major essential elements required for
physiological mechanisms of plant growth. Bio-fertilizers importing a large population of effective microorganisms in the
active field of root system, increase plants power to absorb more nutrients.Continuous cropping with low or no fertilizer
inputs, nutrient losses led to decline in soil fertility and might be develop a stress condition due to which high level of lipid
peroxidation, proline content and increased peroxidise activity. It shows that chemical fertilizer in combinations as well as
chemical fertilizers with biofertilizer reduce the stress in wheat seedlings.

ACKNOWLEDGEMENTS
We express our sincere thanks to DR. K.N. Chaturvedi, Principal and Prof. R.S. Maheshwari, HOD, Biochemistry,
Govt. Holkar Science College (M.P.) for providing necessary laboratory facilities and encouragement.
REFERENCES

1.

Bates L. S., Waldran R. P. and Teare I. D., (1973).Rapid determination of free proline for water stress studies.
Plant soil. 39: 205-208.

2.

Chen J., (2006) - The combined use of chemical and organic fertilizers for crop growth and soil fertility.
International workshop on Sustained Management of the Soil-Rhizosphere System for Efficient Crop Production
and Fertilizer Use 16. 20. p:1-10.

3.

Heath R. L. & Packer L., (1968). Photoperoxidation in isolated chloroplasts. I-Kinetics & stoichiometry of fatty
acids peroxidation. Arch. Biochem. Biophys. 125: 189 -198.

4.

Jithesh M. N., Prashanth S. R., Sivaprakash K. R. and Parida A. K. (2006). Antioxidative response mechanisms in
halophytes: their role in stress defence Journal of Genetics, Vol. 85, No. 3.

5.

Lvai L., Szilvia Veres, Nra Bkonyi, and va Gajdos (2008). Agronomski Glasnic, 3, p: 263-271.

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editor@tjprc.org

206

Sandhya Sharma & Angurbala Bafna

6.

Pimentel D., (1996). Green Revolution and chemical hazards. Sci Total Environ 188: 86-98.

7.

Sharma P., Jha A. B., Dubey R. M., and Pessarakli M., (2012). Article Reactive Oxygen Species, Oxidative
Damage, and Antioxidative Defense Mechanism in Plants under Stressful Conditions Journal of Botany Volume
2012, Article ID 217037, 26 pages.

8.

Summer J. B. and Gjessing F. C., (1943). Arch Biochem. 2:29.

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