Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183

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Biocatalysis and Agricultural Biotechnology

journal homepage: www.elsevier.com/locate/bab

Pre-sowing seed magnetic field treatment influence on germination,

seedling growth and enzymatic activities of melon (Cucumis melo L.)
Munawar Iqbal a,n, Zia ul Haq b, Yasir Jamil c, Jan Nisar d
a
Department of Chemistry, Qurtuba University of Science and Information Technology, Peshawar 25100, KPK, Pakistan
b
Department of Physics, University of Agriculture, Faisalabad 38040, Pakistan
c
Laser Spectroscopy Laboratory, Department of Physics, University of Agriculture, Faisalabad 38040, Pakistan
d
National Centre of Excellence in Physical Chemistry, University of Peshawar, Peshawar 25120, Pakistan

art ic l e i nf o a b s t r a c t

Article history: Pre-sowing seed magnetic field (MF) treatment effects on germination, seedling growth, chlorophyll
Received 17 November 2015 contents and enzymatic activities of a melon cultivar were evaluated. Melon seeds were exposed to MF
Received in revised form strength of 100 mT and 200 mT for 5-20 min under laboratory conditions. MF pre-sowing treatment
12 March 2016
enhanced the melon seed germination (%), root and shoot lengths, vigor indices I and II, seedling fresh
Accepted 2 April 2016
Available online 2 April 2016
and dry masses, leaf area, alpha amylase, protease, catalase, chlorophyll “a” and “b” up to 14.6%, 36.4%,
22.8%, 40.6%, 28.8%, 9.6%, 12.9%, 50.0%, 80.0%, 92.5%, 36.5%, 50.4% and 80.9%, respectively. Moreover, the
Keywords: mean germination time (MGT) was reduced by 6.7% in response to MF pre-sowing seed treatment. Pre-
Magnetic field sowing MF treatment can improve melon germination, seedling growth and related attributes and could
Melon
possibly enhance productivity. In future research the MF pre-sowing seed treatment needs to be tested at
Germination
the field level. If the trials on actual producing farms with full farmer participation are conducted, then it
Seedling growth
Enzymatic activities will help in linking of farmer-based results with the research-based results.
& 2016 Elsevier Ltd. All rights reserved.

1. Introduction during germination and seedling growth (Vashisth and Nagarajan,

2010), accelerated nutrient uptake (Eşitken and Turan, 2004), in-
The application of magnetic field in agricultural sector has been creased chlorophyll contents and leaf area (Iqbal, J., et al., 2013;
emerged as an efficient method to stimulate the plant biological Iqbal, M., et al., 2012a), enabled the seedlings and plants to cope
characteristics and has been extensively studied for assessing the against environmental stresses (Yao et al., 2005), enhanced re-
MF pre-sowing seed treatment effect on germination, growth and generation capability in plant tissues (Telci Kahramanogullari
yield. The technique was found out to be far better than chemical et al., 2012), reduced harmful pathogens effects (De Souza et al.,
fertilizers vis-à-vis productivity and safety. Undoubtedly, MF pre- 2006) and prompted the biochemical reactions (Vashisth and
sowing seed treatment is eco-friendly as well as non-polluting to Nagarajan, 2010) and resultantly, the yield and crop productivity
the soil and environment because clean environmental is also were found to increase many folds (De Souza et al., 2005, 2008;
important after food security (Adesola et al., 2016; Babarinde and Iqbal, M., et al., 2012b; Jamil et al., 2012; Naz et al., 2012).
Onyiaocha, 2016; Gangadhara and Prasad, 2016; Iqbal and Bhatti, Melon (Cucumis melo L.) is an important horticultural crop with
2014, 2015; Iqbal et al., 2012a; Iqbal and Khera, 2015; Jamal et al., a worldwide production of 27.3 million metric tons, with China,
2015; Peter and Chinedu, 2016; Qureshi et al., 2015; Rane et al., Iran, Turkey, Egypt and United States accounting for 68% of the
2015; Sayed, 2015; Ukpaka, 2016; Ukpaka et al., 2015). The use of world production (Sharma et al., 2014). In Pakistan, the Melon
MF pre-sowing seed treatment not only fascinated the scientific (Cucumis melo L.) germination is poor at low temperature (Nerson
community but also got the attention of the farmers as the tech- and Govers, 1986). Optimum seed germination and seedling
nique was observed to be affordable and safe for them (De Souza emergence occur at relatively high temperatures (25–28 °C). Poor
et al., 2014; Iqbal, J., et al., 2013; Iqbal, M., et al., 2012a). It has been germination is a common phenomenon at sub-optimal tempera-
reported that MF pre-sowing seed treatment increased water tures and a major problem faced by the growers. Melon cultivation
uptake in seed (De Souza et al., 2014), enhanced enzymatic activity in late winter and early spring is a big problem in Pakistan. De-
layed and reduced seedling emergence cause non-uniform plant
n
Corresponding author. development, thereby extending melon fruit maturation for early
E-mail address: bosalvee@yahoo.com (M. Iqbal). markets (Farooq et al., 2007). Therefore, there is need to use some

http://dx.doi.org/10.1016/j.bcab.2016.04.001
1878-8181/& 2016 Elsevier Ltd. All rights reserved.
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183 177

methods that might be successful in improving the Melon ger- respectively.

mination. In this regard, the MF pre-sowing seed treatment has
Seed germinated
been proven its effects on germination of various grains and hor- Germination (%) = × 100
total seeds (1)
ticulture crops as well as vegetables (Iqbal, M., et al., 2012a; Jamil
et al., 2012; Naz et al., 2012; Haq et al., 2012). To best of our
Vigor Index I = G (%) × TSL (2)
knowledge, the MF effect on Melon (native to Pakistan) germina-
tion and seedling growth has not been studied previously. This fact
Vigor Index II = G (%) × TSDW (3)
motivated us to investigate the MF pre-sowing Melon seed treat-
ment effect on germination, seedling growth, chlorophyll contents Mean germination time (MGT) was measured as per following
and enzymatic activities during germination and early growth reported method (Flórez et al., 2007) using relation shown in Eq.
stages. (4), where n ¼number of seeds germinated on day D and
D¼ number of days counted from the beginning of germination.
∑ Dn
2. Material and methods MGT =
∑n (4)
Melon (Cucumis melo L., var Ravi) seeds were obtained from Leaf area was measured by portable leaf area meter (Model
Ayub Agricultural Research Institute (AARI), Faisalabad, Pakistan. YMJ-A, China). The root and shoot lengths (cm), seedling fresh and
The experiments were conducted in February, 2013. The climatic dry weights were calculated after 20 days of germination. Dry
conditions recorded from meteorological station, University of weights were determined after drying at 105 °C for 15 min, fol-
Agriculture, Faisalabad. The mean values of temperature, relative lowed by incubation at 80 °C for 24 h (Petropoulou et al., 1995).
humidity, sun shine, rain fall and wind speed were 35.7 °C, 40.5%, The root and shoot lengths were determined by scale from root
8.4 h, 11.5 mm and 8.01 km/h, respectively. neck to tip and shoot from base to tip (Iqbal, J., et al., 2013). All
The MF treatment process was performed in the Department of measurements were performed in triplicate and data thus, ob-
Physics, University of Agriculture, Faisalabad. For MF treatment, tained was averaged.
uniform seeds were selected by hand picking and seeds having For measuring the chlorophyll contents, fresh leaves, 20-day-
visible defects, insect damage or malformation were discarded. old (0.1 g) were ground in the presence of 1 mL acetone and the
The procedure and electromagnet specification have already been volume was made up to 5 mL by adding more acetone. The slurry
reported (Iqbal, J., et al., 2013; Naz et al., 2012). The seeds were was then centrifuged for 5 min. The supernatant was collected and
soaked in water prior to MF treatment for 2 h. After removing the its absorbance was measured at 645 and 663 nm using UV/visible
extra moisture by pressing in filter paper and air drying, seeds spectrophotometer-CE Cecil 7200, UK. The chlorophyll “a” (chl a)
were exposed to MF treatments (Table 1). The MF treated seeds and chlorophyll “b” (chl b) were calculated using relations ((Eqs.
were sown in pots (30
25 cm) filled in by sand and kept under (5)–6)), respectively. Where V¼volume, W¼weight of fresh leaves
control conditions. Before sowing, seeds were sterilized with 0.05% taken and abs¼absorbance (Iqbal, M., et al., 2016; Maqsood et al.,
HgCl2 solution for 10 min and washed with distilled water. Before 2013).
pot filling, the sand was properly washed with distilled water and
saturated with Hoagland's nutrient solution (half-strength). Seeds
⎧ 12. 7 (Abs 663)–2. 69 (Abs 645) ⎫
Chl a=⎨ ⎬×V
were placed on the sand surface and the pots were filled up to the ⎩ 1000 × W ⎭ (5)
upper mark with additional sand layer in such as way that seed
depth remained in the range of  2 cm. For maintenance of ap- ⎧ 22. 9 (Abs 645)–4. 68 (Abs 663) ⎫
Chl b=⎨ ⎬×V
propriate moisture content; the top surface of the sand was ⎩ 1000 × W ⎭ (6)
moistened twice a day using a water sprayer.
The seeds thus germinated were calculated following Associa- The enzymatic catalase (CAT) was determined as per procedure
tion of Official Seed Analysis rules (Carbonell et al., 2000). For reported earlier (Perveen et al., 2011). For protease (PRT) and α-
germination percentage estimation, emerged seedlings (cotyle- amylase activity measurement, the method is reported elsewhere
dons visible at the sand surface) were counted daily until further (Jamil et al., 2013). To measure PRT and α-amylase activity, seeds
germination of the seeds ceased. The germination percentage was were grown in Petri plates in laboratory immediately after ger-
estimated using relation (Eq. (1)). The Vigor indices I and II were mination, whereas CAT activity was measured in 20-day-old. For
calculated using relations ((Eqs. (2)–3)) (Vashisth and Nagarajan, the extraction of CAT, fresh leaves (20-day-old) (0.5 g) were
2010), where G is germination percentage and TSL & TSDW are ground in 5 mL of 50 mM cooled phosphate buffer (pH 7.8) using a
total seedling length (TSL) and total seedling dry weight (TSDW), pestle and mortar. After filtration through a cheese-cloth, the
homogenate was centrifuged for 20 min at 4 °C and the super-
Table 1 natant was used for enzyme assays (Perveen et al., 2011). Analysis
The MF dose (strength-mT and exposure time-min) and experiment layout used to of variance of the data for each attribute was computed using
design experiment under randomized complete block design. statistical program STATISTICA (Version 8.1). The least significance
difference test at 5% level of probability was applied to test the
Treatments MF Time Experimental design
differences among means.
mT min Replicate I Replicate II Replicate III

T0 0 0 T0R1 T0R2 T0R3 3. Results and discussion

T1 100 5 T1R1 T1R2 T1R3
T2 200 5 T2R1 T2R2 T2R3
T3 100 10 T3R1 T3R2 T3R3 In present investigation, Melon seed germination, seedling
T4 200 10 T4R1 T1R2 T1R3 growth, chlorophyll contents, leaf area and enzymatic activities
T5 100 15 T5R1 T5R2 T5R3 were studied under the effect of MF pre-sowing seed treatment. It
T6 200 15 T6R1 T6R2 T6R3 was observed that the MF pre-sowing Melon seed treatment
T7 100 20 T7R1 T7R2 T7R3
T8 200 20 T8R1 T8R2 T8R3
augmented the germination, growth and enzymatic activities
considerably.
178 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183

96 2166.3, 2033.1, 2314, 2393.3, 2393.3, 2497.8 and 2358.4, respec-

(A)
g tively for T1, T2, T3, T4, T5, T6, T7, and T8, while it was 1920.0 in
94
fg control (Fig. 2(B)). The percentage increments in vigor index I over
92 f control were 0.19%, 10.5%, 7.4%, 19.3%, 22.7%, 19.5%, 25.9% and
de 40.6% and for vigor index II, these values were 4.4%, 12.26%, 5.6%,
90 cd 19.6%, 23.6%, 23.6%, 28.8% and 21.8% in comparison to control for
c
the treatments T1, T2, T3, T4, T5, T6, T7, and T8, respectively (Table 2).
Germination (%)

88
The root and shoot lengths were also enhanced noticeably in re-
86 sponse to MF pre-sowing Melon seed treatment and T1, T2, T3, T4,
T5, T6, T7, and T8 showed the root lengths of 3.3, 3.3, 3.5, 3.7, 3.8,
84 b 3.4, 3.2 and 3.5 (cm), respectively, while root length in control was
a a 3.3 cm (Fig. 3(A)). The percentage effects over control for T1, T2, T3,
82
T4, T5, T6, T7, and T8 were 0.0%, 0.0%, 6.1%, 12.1%, 15.2%, 23.0%, 3.0%
80 and 36.4%, respectively (Table 2). The shoot lengths of 5.6, 5.6, 5.7,
5.9, 5.8, 5.8, 6.0, 6.5 and 6.8 (cm) were observed in T1, T2, T3, T4, T5,
78 T6, T7, and T8, respectively in treated group (Fig. 3(B)). The percent
increase in shoot lengths were observed to be 0.0%, 2.5%, 6.1%,
76
4.3%, 3.6%, 8.1%, 16.9% and 22.8% in case of T1, T2, T3, T4, T5, T6, T7,
T0 T1 T2 T3 T4 T5 T6 T7 T8
and T8, respectively as compared to control (Table 2). Previous
Treatments studies have also revealed that the MF treatment had promising
(B) effects on germination and seedling growth of seeds correspond-
a ing to different plants (Anand et al., 2012; Flórez et al., 2007) and
6.1
similar trend was observed in present investigation. Seedling fresh
and dry weights were also increased in response to MF treatment.
6.0 d e The seedling (root and shoot) fresh weights were observed to be
de 520.0, 530.0, 515.0, 540.0, 550.0, 535.0, 530.0 and 540.0 (g) for T1,
e
5.9 cde T2, T3, T4, T5, T6, T7, and T8, respectively in 20-day-old seedlings,
whereas it was noted to be 500.0 in control (Fig. 4(A)). The seed-
MET (days)

cd ling dry weights were 24.5, 24.9, 25.1, 26.0, 26.3, 26.5, 27.2 and
5.8 bc
26.8 (g) in case of T1, T2, T3, T4, T5, T6, T7, and T8 treatments, re-
spectively and in control seedling dry weight was 24 g (Fig. 4(B)).
5.7 b
The percentage increments in seedling fresh weights were 3.9%,
5.7%, 2.9%, 7.7%, 9.6%, 6.7%, 5.8% and 7.7% for T1, T2, T3, T4, T5, T6, T7,
5.6 and T8 treatments, respectively (Table 2). The seedling dry weights
increased up to 2.0%, 3.7%, 4.5%, 8.2%, 9.4%, 10.2%, 12.9% and 11.4%
for T1, T2, T3, T4, T5, T6, T7, and T8 treatments, respectively versus
5.5
control. The leaf area of Melon seedlings, raised from magnetically
treated seeds also enhanced (Fig. 4(C)) and 30.6%, 16.7%, 32.2%,
T0 T1 T2 T3 T4 T5 T6 T7 T8
5.6%, 50.0%, 19.4%, 45.8% and 44.4% higher leaf areas were recorded
Treatments in seedlings, raised from T1, T2, T3, T4, T5, T6, T7, and T8 treated
Fig. 1. Melon seed pre-sowing magnetic field treatment effect on germination seeds, respectively (Table 2). These findings were comparable with
percentage (A) and mean emergence time (MET) (B). [T0 (control), T1 (100 mT, earlier studies that MF influenced the plant growth and develop-
5 min), T2 (200 mT, 5 min), T3 (100 mT, 10 min), T4 (200 mT, 10 min), T5 (100 mT, ment at lateral stages (Iqbal, J., et al., 2013; Iqbal, M., et al., 2012b;
15 min), T6 (200 mT, 15 min), T7 (100 mT, 20 min), and T8 (200 mT, 20 min)]. Data
Jamil et al., 2012). MF doses of 125 mT and 250 mT revealed early
are representative of three independent experiments and values are expressed in
mean 7SD. Different superscript letters following the means indicate a significant growth in maize plant (Flórez et al., 2007), higher shoot length in
difference (Pr 0.05). MF treated pea plant is observed (Iqbal, M., et al., 2012a), eight-
days-old chick-pea plant showed 2.67 cm difference in shoot
length in comparison to control (Nasher, 2008) and MF increased
The germination data revealed that MF pre-sowing seed root length of sunflower plant significantly (P o0.01) (Vashisth
treatment enhanced germination percentage significantly. The and Nagarajan, 2010). Similarly, the MF also enhanced leaf area
treatment T1, T2, T3, T4, T5, T6, T7, and T8 showed the germination and seedling growth in 15-days-old Lentil plants in response to
percentage of Melon seed up to 82, 87, 81, 89, 91, 90, 92 and 88 (%), 0.06–0.36 T MF treatment for 5, 10, 20 min (Shabrangi et al., 2013),
respectively, whereas it was 80% in control (Fig. 1(A)). The treat- number of leaves in barley plant raised from magnetically treated
ment T1, T2, T3, T4, T5, T6, T7, and T8 showed germination percen- seed also increased (Carbonell et al., 2000). Similarly, De Souza
tages of 2.4%, 8.5%, 1.2%, 11.0%, 13.4%, 12.2%, 14.6% and 9.8% higher et al. (2014) recorded higher plant height in onion as a result of
as compared to control, respectively (Table 2). The MET reduced extremely low frequency non-uniform magnetic field treatment
considerably when seed were exposed to different MF dose. The and broad bean MF treated seed showed early seedling growth in
MET values of 5.9, 5.9, 5.8, 5.9, 5.8, 5.9, 5.7 and 5.6 (days) were comparison to control (Rajendra et al., 2005).
recorded for T1, T2, T3, T4, T5, T6, T7, and T8 ‘s, respectively, whereas The chlorophyll “a” and “b” contents were also increased as a
it was 6 days in control (T0) (Fig. 1(B)). The percentage reductions result of MF treatment (Fig. 5(A) and (B)). The chlorophyll “a” in-
in MET values for Melon seed for T1, T2, T3, T4, T5, T6, T7, and T8 were creased up to 50.7%, 23.1%, 37.1%, 21.7%, 44.8%, 46.2%, 47.6% and
1.67%,  2.50%,  3.34%,  1.67%,  4.17%, 2.17%,  5.00% and 50.4% as compared to control for T1, T2, T3, T4, T5, T6, T7, and T8
6.67%, respectively versus control (Table 2). The vigor index I treatments, respectively, whereas chlorophyll “b” showed the
values were recorded to be 710.1, 783.0, 761.4, 845.5, 870.0, 846.9, percentage increments of 50.7%, 19.2%, 31.9%, 17.0%, 76.6%, 61.7%,
892.4 and 997.0 for T1, T2, T3, T4, T5, T6, T7, and T8, respectively 68.1% and 80.9% versus control for the treatments T1, T2, T3, T4, T5,
(Fig. 2(A)), whereas vigor index II values were observed as 2009, T6, T7, and T8, respectively (Table 2). The leaf area increased up to
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183 179

Table 2
Percentage effect of MF pre-sowing seed treatment on germination, seedling growth and enzymatic activities.

Parameters T0 T1 T2 T3 T4 T5 T6 T7 T8

Germination (%) 0.0a 2.45b 8.5c 1.2a 11.0de 13.4fg 12.2ef 14.6g 9.8cd
Root length (cm) 0.0a 0.0a 0.0a 22.1c 30.1e 36.4f 14.0b 15.3b 26.4c
Shoot length (cm) 0.0a 0.0a 2.5b 6.1d 4.3c 3.6c 8.1e 16.9f 22.8g
Vigor index I 0.0a 0.2a 10.5c 7.4b 19.3d 22.7e 19.5d 25.9f 40.6g
Vigor index II 0.0a 4.4b 12.3c 5.6b 19.6d 23.6e 23.6e 28.8f 21.8de
MET (days) 0.0a  1.7d  2.5de  3.3cde  1.7e  4.2cd  2.2e  5.0BCE  6.7b
Seedling fresh weight (g) 0.0a 3.9b 5.7c 2.9b 7.7d 9.6e 6.7cd 5.8c 7.7d
Seedling dry weight (g) 0.0a 2.0b 3.7c 4.5c 8.2d 9.4cd 10.2de 12.9g 11.4fg
Leaf area (cm2) 0.0a 30.6d 16.7c 32.2d 5. 6b 50.0g 19.4c 45.8ef 44.4e
Alpha amylase (mg  g f. wt) 0.0a 50.7b 40.0c 53.3c 66.7d 80.0e 63.3d 53.3c 66.7d
Protease (mg  g f. wt) 0.0a 50.7b 34.2c 42.5d 59.2e 67.5f 17.5e 92.5g 85.8g
Catalase (U mg  1 protein 0.0a 18.5b 21.2c 27.0de 30.8e 21.2c 25.0d 34.6f 36.5f
Chlorophyll “a” (mg/g) 0.0a 19.5b 23.1c 37.1d 21.7c 44.8e 46.2e 47.6e 50.4e
Chlorophyll “b” (mg/g) 0.0a 14.6a 19.2b 31.9c 17.0b 76.6e 61.7d 68.1d 80.9e

[T0 (control, for percentage effect calculation, the control effect was considered 0%), T1 (100 mT, 5 min), T2 (200 mT, 5 min), T3 (100 mT, 10 min), T4 (200 mT, 10 min), T5
(100 mT, 15 min), T6 (200 mT, 15 min), T7 (100 mT, 20 min), and T8 (200 mT, 20 min)]. Data are representative of three independent experiments. Different superscript letters
following the means indicate a significant difference (Pr 0.05).

1050 (A)
(A) f
g
3.8 e
1000

950
f c
Root length (cm)
3.6
900 e c
Vigor index I

d d
850 b b

c 3.4 a
800 a a
b
750 a
a
3.2
700

650
T0 T1 T2 T3 T4 T5 T6 T7 T8
T0 T1 T2 T3 T4 T5 T6 T7 T8
Treatments
Treatments
7.2
(B) f (B)
2600
7.0
g
2500 e e
de 6.8
d f
2400
6.6
Shoot length (cm)

2300
Vigor index II

c 6.4

2200 6.2 e
b d
2100 6.0 c
b c
a b
2000 5.8 a
a
1900 5.6

5.4
1800
5.2
T0 T1 T2 T3 T4 T5 T6 T7 T8
T0 T1 T2 T3 T4 T5 T6 T7 T8
Treatments Treatments
Fig. 2. Melon seed pre-sowing magnetic field treatment effect on vigor index I
Fig. 3. (A) Root and (B) shoot lengths of Melon, raised from magnetically treated
(A) and vigor index II (B) (explanation as in Fig. 1).
seeds (explanation as in Fig. 1).

30.6%, 16.7%, 32.2%, 5.6%, 50.0%, 19.4%, 45.8% and 44.4% for T1, T2, positive effect on chlorophyll contents and related physiological
T3, T4, T5, T6, T7, and T8 treatments, respectively (Fig. 5(C), Table 2). attributes i.e., cucumber seed, treated at 0.2 and 0.45 T MF showed
These findings are in line with previous studies that MF had significant change in leaf chlorophyll contents (Yao et al., 2005).
180 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183

(A) 2.0
(A) e
600
c e d
b d cd c e e
a b e
1.9
500

1.8
d
Seedling fresh weight (g)

400
1.7

Chl a (mg/g)
300
1.6 c c
200 1.5 b

100 1.4

1.3
0 a
1.2
-100
T0 T1 T2 T3 T4 T5 T6 T7 T8 T0 T1 T2 T3 T4 T5 T6 T7 T8
Treatments Treatments

28.0
(B) (B)
g 0.85 e e
27.5 fg
de 0.80 d
27.0
cd 0.75 d
26.5
Seedling dry weight (g)

d 0.70
26.0
0.65
Chl b (mg/g)

c c
25.5 0.60
c
25.0 b 0.55 b b
a a a
24.5 0.50

24.0 0.45

0.40
23.5
0.35
23.0
T0 T1 T2 T3 T4 T5 T6 T7 T8 T0 T1 T2 T3 T4 T5 T6 T7 T8
Treatments Treatments
Fig. 4. Effect of magnetic field pre-sowing seed treatment effect on seedling fresh
weight (A) and seedling dry weight (B) (explanation as in Fig. 1).
4.6
(C)
g
4.4
Similarly, higher chlorophyll contents in pea plant raised from e
4.2 e
magnetically treated seeds has also been reported (Iqbal, M., et al.,
2012a). In another study, MF dose of 100 mT for 10 min and 4.0
170 mT for 3 min enhanced leaf area in tomato plant and 160 mT 3.8 d
d
Leaf area (cm2)

for 15 and 20 min in onion also affected leaf area positively (De
3.6
Souza et al., 2014, 2008).
Germination and seedling emergence demand high energy for 3.4
c
respiration of the seed embryo and cotyledon. The mobilization of c
3.2
seed storage proteins represents one of the most important post-
germination events in the growth and development of the seed- 3.0
ling. Proteolytic enzymes play a central role in the biochemical b
2.8
mechanism of germination (Vashisth and Nagarajan, 2010). The
enzymes are involved in the germination process and metabolism 2.6 a
i.e., amylases and proteases are responsible for breakdown of food 2.4
reservoir in seed to provide nutrition to newly germinated plu-
mule and radical, whereas catalase is a antioxidant enzyme and T0 T1 T2 T3 T4 T5 T6 T7 T8
defends seedling from the environmental stress during growth Treatments
process and in this way enzymes play a key role in the germination
Fig. 5. (A) Chlorophyll “a”, (B) Chlorophyll “b” (C) leaf area of Melon seedlings,
and seedling growth (Jamil et al., 2013; Perveen et al., 2011). The
raised from magnetically treated seeds (explanation as in Fig. 1).
enzymes activities measured during germination and later stages
revealed that the enzymatic activities were considerably higher in
seedlings, raised from magnetically treated seeds. The increased
activities of these enzymes are beneficial for rapid and uniform
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183 181

2.8 germination and hence better crop establishment. The alpha

(A)
amylase activities were 1.5, 1.9, 2.1, 2.3, 2.5, 2.3, 2.1 and 2.3 (mg/g
2.6 e fresh wt) for T1, T2, T3, T4, T5, T6, T7, and T8 treatments, respectively
and in control it was 1.3 mg/g (Fig. 6(A)) and percentage increase
Alpha amylase (mg-g fresh wt)

2.4 d d in alpha amylase activities were 50.7%, 40.00%, 53.33%, 66.67%,

d
80.00%, 63.33%, 53.33% and 66.67% in comparison to control (Ta-
2.2 c c ble 2). Similarly, protease activity was also higher in treated seeds
and activities of 1.2, 1.4, 1.5, 1.7, 1.8, 1.2, 2.1 and 2.0 (mg/g fresh wt)
2.0 c were observed in T1, T2, T3, T4, T5, T6, T7, and T8 treated seeds, re-
spectively (Fig. 6(B)). The percentage increase in protease activities
1.8
were 50.7%, 34.2%, 42.5%, 59.2%, 67.5%, 17.5%, 92.5% and 85.8% for
b T1, T2, T3, T4, T5, T6, T7, and T8, respectively over control and 50.7%,
1.6
21.2%, 26.9%, 30.8%, 21.2%, 25.0%, 34.6% and 36.5% increment in
catalase activity was recorded for T1, T2, T3, T4, T5, T6, T7, and T8,
1.4 a
respectively (Fig. 6(C)) and (Table 2). Previous studies also support
1.2
these findings that MF may enhance the enzymatic activity in MF
treated seeds and seedlings at lateral stages of development i.e.,
T0 T1 T2 T3 T4 T5 T6 T7 T8 enhanced enzymatic activities in sunflower seeds exposed to static
magnetic field has been reported (Vashisth and Nagarajan, 2010),
Treatments
extremely low frequency MF exposure enhanced the enzymatic
activities in Zea mays (Shabrangi et al., 2013). MF pre-sowing
(B)
2.2 treatment affected enzyme activity in the sea cucumber (Tang
g
et al., 2015) and in germinating seeds of broad bean (Rajendra
g
et al., 2005).
2.0
The MF pre-sowing seed treatment stimulated the germination,
f
seedling growth (fresh and dry, lengths), chlorophyll contents, leaf
Protease (mg-g fresh wt)

1.8 e e area and enzymatic activities of Melon. Results showed that the
germination and other moieties can be enhanced by applying an
1.6 d appropriate combination of MF strength for suitable exposure
c time. It was also observed that the germination, seedling growth,
1.4 chlorophyll contents and enzyme characteristics amplified at cer-
b tain levels of MF dose and exposure time i.e., 100 mT for 15 and
1.2 20 min and 200 mT for 10 and 15 min were found more effective
as compared to other levels of MF dose and exposure time. The MF
a
1.0
action mechanisms on seed germination and seedling growth is
not well known, however, several theories have been proposed,
including biochemical changes or altered enzyme activities. Seed
0.8
germination stimulation might be attributed to a combined effect
T0 T1 T2 T3 T4 T5 T6 T7 T8
of biochemical, physiological, metabolic and enhanced enzymatic
Treatments action. It is considered that MF alters cell membrane permeability
and transfer water and energy signals into the cell (Reina et al.,
(C) f 2001) and resultantly, metabolic pathways may influence (Iqbal,
65 f
M., et al., 2013). The enzymes which are necessary for seed ger-
e mination at particular stages of germination were found higher in
de magnetically treated seeds during seed germination (Vashisth and
60 d Nagarajan, 2010). Another theory stated that MF affects the bio-
CAT (U mg-1 protein)

c c logical objects by non-conventional spins, free radicals, liquids

crystals or mobile electron charges. Chemically these free radicals
55 are very active species and are responsible for biochemical and
b
physiological processes during seed germination (Podleoeny et al.,
2004). The increased water absorption has also been reported
50
which might be responsible for enhanced germination (Reina
a et al., 2001), especially in case of bitter gourd since thick coat is
one of the reasons for low germination. The enhanced growth and
45
development and growth of bitter gourd at lateral stages of de-
velopment might be attributed to the a-amylase, dehydrogenase
and protease activities (Vashisth and Nagarajan, 2010), because
40
enzymes are responsible for the degradation of food reserves
T0 T1 T2 T3 T4 T5 T6 T7 T8
stored in seed during germination and seedling growth (Iqbal, M.,
Treatments et al., 2012a). The enhanced leaf area in seedlings raised from
Fig. 6. Effect of magnetic field pre-sowing seed treatment on (A) Alpha amylase magnetically treated seeds may result in a greater interception of
(mg/g fresh wt), (B) Protease (mg/g fresh wt) activities in Melon germinating seeds light and might be responsible for higher chlorophyll contents and
and (C) Catalase (U/mg protein) activity in Melon 20-day-old seedlings (explana- photosynthetic rates since photosynthetic rate and influx of water
tion as in Fig. 1).
as a result of magnetic treatment is reported (Hoff, 1981). The
enhanced chlorophyll contents might be due to the intake of ions
responsible for photosynthetic rate. It has been reported that MF
182 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 6 (2016) 176–183

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be enhanced and previous finding also correlate well with this phys. 14, 599–665.
hypothesis i.e., pre-sowing magnetic treatments of tomato seeds Iqbal, J., Cecil, F., Ahmad, K., Iqbal, M., Mushtaq, M., Naeem, M., Bokhari, T., 2013.
increase the growth and yield of plants (De Souza et al., 2006, Kinetic study of Cr (III) and Cr (VI) biosorption using Rosa damascena phy-
tomass: a rose waste biomass. Asian J. Chem. 25, 2099–2103.
2005), pea (Podleoeny et al., 2005), lettuce (De Souza et al., 2008) Iqbal, M., Ahmad, I., Hussain, S.M., Khera, R.A., Bokhari, T.H., Shehzad, M.A., 2013.
and okra (Naz et al., 2012). This increased yield in turn accredited Optimization of pre-sowing magnetic field doses through RSM in pea. Int.
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Iqbal, M., Khera, R.A., 2015. Adsorption of copper and lead in single and binary
The effect of pre-sowing MF seed treatment on melon (Cucumis metal system onto Fumaria indica biomass. Chem. Int. 1, 157b–163b.
Iqbal, M., Muhammad, D., Zia-ul-Haq, Jamil, Y., Ahmad, M.R., 2012b. Effect of pre-
melo L.) biological attributes was evaluated and as a MF pre- sowing magnetic field treatment to garden pea (Pisum sativum L.) seed on
sowing seed treatment, melon seed germination characteristics, germination and seedling growth. Pak. J. Bot. 44, 1851–1856.
seedling growth, chlorophyll contents and enzymatic activities Iqbal, M., ul Haq, Z., Malik, A., Ayoub, C.M., Jamil, Y., Nisar, J., 2016. Pre-sowing seed
magnetic field stimulation: a good option to enhance bitter gourd germination,
increased significantly as compared to control. The MF dose seedling growth and yield characteristics. Biocat. Agric. Biotechnol. 5, 30–37.
(strength and seed exposure time) differentially enhanced germi- Jamal, M.A., Muneer, M., Iqbal, M., 2015. Photo-degradation of monoazo dye blue 13
nation, seedling growth, enzymatic activities and biosynthesis of using advanced oxidation process. Chem. Int. 1, 12–16.
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chlorophyll. Generally, low strength/longer exposure and vice mushroom using magnetic field treatment. Int. Agrophys. 26, 375–380.
versa were found promising. The use of pre-sowing MF seed Jamil, Y., Perveen, R., Ashraf, M., Ali, Q., Iqbal, M., Ahmad, M.R., 2013. He–Ne laser-
treatment is suggested for enhancing germination and seedling induced changes in germination, thermodynamic parameters, internal energy,
enzyme activities and physiological attributes of wheat during germination and
growth in melon. Future studies should focus on yield character-
early growth. Laser Phys. Lett. 10, 045606.
istics and trials should be done on the farm level with participa- Maqsood, M., Shehzad, M.A., Ali, S.N.A., Iqbal, M., 2013. Rice cultures and nitrogen
tion of farmers to link farmer-based evaluation of results with the rate effects on yield and quality of rice (Oryza sativa L.). Turk. J. Agric. 37,
lab-based results. 665–673.
Nasher, S.H., 2008. The effect of magnetic water on growth of chick-pea seeds. Eng.
Technol. 26, 16–20.
Naz, A., Jamil, Y., Iqbal, M., Ahmad, M.R., Ashraf, M.I., Ahmad, R., 2012. Enhancement
Acknowledgments in the germination, growth and yield of okra (Abelmoschus esculentus) using
pre-sowing magnetic treatment of seeds. Indian J. Biochem. Biophys. 49,
211–214.
Ayyub Agriculture Research Institute is acknowledged for pro- Nerson, H., Govers, A., 1986. Salt priming of muskmelon seeds for low temperature
viding Melon seed. germination. Sci. Hort. 28, 85–91.
Perveen, R., Jamil, Y., Ashraf, M., Ali, Q., Iqbal, M., Ahmad, M.R., 2011. He-Ne Laser-
Induced improvement in biochemical, physiological, growth and yield char-
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