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Original Article
FTIR and multivariate analysis to study the effect of bulk and nano
copper oxide on peanut plant leaves
S. Suresh a, b, *, S. Karthikeyan c, K. Jayamoorthy d
a
Research and Development Center, Bharathiar University, Coimbatore 641 046, Tamilnadu, India
Department of Physics, St. Joseph's College of Engineering, Chennai 600 119, Tamilnadu, India
c
Department of Physics, Dr. Ambedkar Government Arts College, Chennai 600 039, Tamilnadu, India
d
Department of Chemistry, St. Joseph's College of Engineering, Chennai 600 119, Tamilnadu, India
b
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 26 July 2016
Accepted 12 August 2016
Available online 22 August 2016
In this article the potential variation in biochemical constituents of peanut plant leaves affect by
presoaking peanut seeds in copper oxide nanoparticles suspension has been studied and compared
with its bulk counterpart. The synthesized nanoparticles were characterized by x-ray diffraction (XRD),
scanning electron microscope (SEM) and transmission electron microscope (TEM) studies. The Fourier
transform infrared (FTIR) analysis shows the most prominent peaks at ~2923 cm1, ~1636 cm1 and
~1033 cm1, which correspond to lipids, protein and carbohydrate content in leaf samples respectively.
The calculated mean ratio of the peak intensities for various frequency regions and total band area
calculation for various band regions explain the variation in lipid, protein and carbohydrate content of
leaf samples. Further the FTIR spectra were processed by de-convolution and curve tting to quantitatively examine the chemical contents and structure changing of the secondary structure of protein.
The calculated integrated band area of b e sheet, b e turn and a e helix secondary structure of protein
varies to greater extent in all samples compared to control. Principal component analysis (PCA) has
been carried out to explain the total variance in secondary structure of protein content in peanut plant
leaves. Principal component 1 (PC1) accounts for 63.50% variation in secondary structure of protein
whereas principal component 2 (PC2) accounts for 29.56%. The application of nanoparticles via presoaking method implies potential variation in biochemical constituents but doesn't affect the growth
of plants considerably.
2016 The Authors. Publishing services by Elsevier B.V. on behalf of Vietnam National University, Hanoi.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords:
Peanut plant
Copper oxide nanoparticles
FTIR
Multivariate analysis
Presoaking method
1. Introduction
Heavy metals are toxic to living organisms at elevated concentrations but it is more essential for many biochemical reactions. The
elements essential for plants are classied as macronutrients (C, H,
O, N, P, K, Ca, Mg, S) and micronutrients or trace elements (B, Cu, Fe,
Mn, Mo and Zn). Lead and cadmium are the non-essential elements
for any biochemical reaction or processes [1e3]. The micronutrients or trace elements are required for plants nutrition at
micro level concentration but at higher concentrations affect the
plants growth badly. Even though macronutrient required for plant
growth, it becomes toxic when the concentration exceeds the
* Corresponding author. Department of Physics, St. Joseph's College of Engineering, Chennai 600 119, Tamilnadu, India.
E-mail address: profsuresh1@gmail.com (S. Suresh).
Peer review under responsibility of Vietnam National University, Hanoi.
http://dx.doi.org/10.1016/j.jsamd.2016.08.004
2468-2179/ 2016 The Authors. Publishing services by Elsevier B.V. on behalf of Vietnam National University, Hanoi. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
344
S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
All the statistical analysis was performed using SPSS 16.0 software. Pearson's correlation matrix was calculated for secondary
structure protein and metal constituent variation in leaf samples.
Principle component analysis was carried out to nd the factors
which inuence the variation in protein content among the leaf
samples treated with bulk and nano copper oxide in comparison
with control sample. Graphical work was carried out using Origin
software 8.0.
3. Results and discussion
3.1. XRD, SEM and TEM analyses of copper oxide nanoparticles
Fig. 1 visualizes the XRD pattern of copper oxide. The formation
of copper oxide phase in 400 C annealed sample was conrmed by
an X-ray diffractometer. The average grain sizes were determined
from the XRD patterns using the Debye-scherrer formula.
0:9*l
b cos q
S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
345
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S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
Table 1
Tentative frequency assignment of FTIR spectra for control, L1, L2, L3 & L4 leaf samples.
Control
3415 (vs)
3347 (vs)
2955 (m)
2921 (vs)
2851 (m)
1736 (w)
1725 (w)
1630 (vs)
1547 (m)
1426 (m)
1383 (m)
1255 (w)
1156 (m)
1105(m)
1070 (m)
1061 (m)
1033 (m)
962 (vw)
L1
L2
L3
L4
3404 (vs)
3384 (vs)
2955 (w)
2921 (m)
2850 (w)
1734 (w)
1717 (m)
1636 (vs)
1541 (m)
1419 (m)
1386 (m)
1247 (w)
1152 (m)
1102(s)
1069 (s)
1061 (s)
1034 (s)
e
3420 (vs)
3365 (vs)
2956 (m)
2920 (s)
2850 (m)
1734 (w)
1717 (m)
1635 (vs)
1540 (s)
1418 (m)
1385 (m)
1260 (w)
1151 (w)
1101(m)
e
1047 (m)
1023 (m)
e
3404 (vs)
3366 (vs)
2956 (m)
2919 (m)
2850 (w)
1735 (w)
1719 (w)
1639 (vs)
1535 (m)
1425 (m)
1384 (m)
1251 (w)
1152 (m)
1102(m)
e
1062 (s)
1035 (s)
e
3404 (vs)
3365 (vs)
2956 (m)
2920 (m)
2850 (w)
1734 (w)
1717 (w)
1636 (vs)
1541 (m)
1419 (m)
1385 (m)
1248 (w)
1153 (m)
1103(m)
1071 (s)
1062 (s)
1034 (s)
e
Table 2
The total band area calculated for control, L1, L2, L3 & L4 leaf samples.
Spectral region (cm1)
Control
L1
L2
L3
L4
3200e3450
409.92 2.084
3000e2800
145.64 0.841
1800e1500
135.35 0.515
1300e1450
66.63 0.478
1000e1200
96.02 1.056
437.74 0.150
6.79
70.17 0.248
51.82
141.66 0.481
4.67
65.21 0.589
2.12
125.23 0.840
30.42
373.94 0.864
8.78
155.61 0.341
6.85
152.81 1.015
12.91
62.72 0.255
5.86
100.38 0.634
4.54
452.92 1.026
10.49
116.79 0.898
19.81
140.78 0.980
4.01
72.30 0.625
8.52
108.61 0.756
13.12
456.23 2.052
11.30
108.37 0.414
25.58
157.20 0.180
16.14
65.93 0.473
1.05
105.26 0.314
9.63
The values are the mean S.E for each group (n 9). Comparisons were done by student t-test. The degree of signicance was p < 0.05.
& L4 samples but slightly increases in L3 by 8.52%. The carbohydrates in all leaf samples found to be increased compared to control
and highest variation was found in L1 sample by 30.42% compared
to control sample. On the whole the total band area calculations
S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
347
Table 3
Mean ratio of peak intensities of the bands at different wave numbers.
Ratio of bands
Control
L1
I1540/I3347
0.5345 0.001
I2954/I2850
1.0096 0.002
I1540/I1635
0.6219 0.008
I1071/I1540
1.0491 0.004
I1734/I1430
0.6535 0.007
I1734/I1540
0.6178 0.003
I1430/I1540
0.9453 0.002
0.4816
9.89
1.2948
28.25
0.5350
13.97
1.4740
40.50
0.6386
6.06
0.6375
3.20
1.0386
9.86
L2
0.002
0.011
0.002
0.004
0.009
0.004
0.007
0.6937
29.79
1.0647
5.45
0.7159
15.11
0.8411
19.83
0.7570
15.84
0.5725
7.32
0.7563
19.99
L3
0.003
0.006
0.005
0.002
0.008
0.003
0.001
0.4336
18.88
1.1745
16.330
0.4868
21.73
1.4688
40.00
0.5640
13.68
0.6414
3.83
1.1372
20.29
L4
0.003
0.008
0.002
0.007
0.005
0.012
0.009
The values are the mean S.E for each group (n 9). Comparisons were done by student t-test. The degree of signicance was p < 0.05.
0.5390
0.85
1.2254
21.37
0.5552
10.73
1.1436
9.01
0.6603
1.05
0.6289
1.80
0.9523
0.74
0.005
0.009
0.001
0.003
0.006
0.003
0.006
348
S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
Table 4
Frequency assignment for secondary protein obtained by self deconvoluted spectra.
Wavenumber
(cm1)
Control
L1
L2
1633 (b e sheet)
1.4785 0.002
2.2533 0.004
1656 (a e helix)
1.0347 0.006
1666 (b e turn)
1.3915 0.006
1673 (b e turn)
0.9997 0.008
1684 (b e turn)
2.1776 0.007
1.4978
1.31
3.7506
66.45
3.1078
200.37
3.6970
165.69
1.9707
97.14
2.7620
26.84
1.7784
20.29
2.7740
23.11
1.7113
65.40
1.6319
17.28
3.1976
219.87
2.6755
22.87
0.005
0.008
0.001
0.002
0.006
0.009
L3
L4
0.8674
41.33
2.4114
7.02
2.3329
125.48
0.8500
38.92
1.6977
69.82
2.2840
4.89
0.002
0.005
0.004
0.006
0.006
0.003
0.004
0.007
0.003
0.007
0.004
0.009
0.8926
39.63
1.9635
12.86
2.7327
164.11
2.1345
53.40
1.9068
90.74
3.6277
66.59
0.004
0.002
0.004
0.001
0.002
0.003
The values are the mean S.E for each group (n 9). Comparisons were done by student t-test. The degree of signicance was p < 0.05.
Table 6
Variation of metal concentration in leaf samples of control, L1, L2, L3 and L4 is
calculated using varimax rotated factor analysis of principal component extraction
method.
Component
1
2
% of variance
Cumulative %
3.175
1.478
63.496
29.555
63.496
93.051
Control
L1
L2
L3
L4
Component
1
0.695
0.980
0.889
0.091
0.966
0.700
0.057
0.015
0.988
0.087
decreasing amounts of residual variance using two factors (varimax rotation) for the samples Control, L1, L2, L3 and L4 of leaf
samples. The main factor (>0.6) for L1, L2 and L4 is noted as factor 1.
Factor 2 contributes control & L3. Factor analysis or principle
component analysis is a useful tool in the examination of multivariate data (Fig. 5). The cumulative percentage of explained
variance in secondary structure of protein of all leaf samples is
93.051%. Factor 1 account for 63.496% of the total data variance so
the variation of secondary structure of protein was established by
accounting the L1, L2 and L4 samples. Factor 2 accounts for 29.555%
which includes the control and L3 samples. Fig. 6 shows the PC
loadings plot for the spectral range 1600e1700 cm1 which
clearly shows the PC1 factor might establish the maximum variation of the secondary structure of protein in leaf samples
compared to other component.
4. Conclusions
Table 5
Pearson correlation matrix for variation in secondary structure protein of control, L1,
L2, L3 & L4 leaf samples.
Wavenumber (cm1)
1633
1648
1656
1666
1673
1684
1633
1648
1656
1666
1673
1684
1
0.709
0.968a
0.927b
0.926b
0.941b
0.709
1
0.853
0.712
0.888b
0.814
0.968a
0.853
1
0.945b
0.990a
0.981a
0.927b
0.712
0.945b
1
0.950b
0.987a
0.926b
0.888b
0.990a
0.950b
1
0.987a
0.941b
0.814
0.981a
0.987a
0.987a
1
a
b
S. Suresh et al. / Journal of Science: Advanced Materials and Devices 1 (2016) 343e350
349
Fig. 5. Principle component analysis explained variance in secondary structure protein of control, L1, L2, L3 & L4 leaf samples.
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