Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37

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Biocatalysis and Agricultural Biotechnology

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

Pre-sowing seed magnetic field stimulation: A good option to enhance

bitter gourd germination, seedling growth and yield characteristics
Munawar Iqbal a,n, Zia ul Haq b, Amna Malik b, Ch. Muhammad Ayoub c, Yasir Jamil d,
Jan Nisar a
a
National Centre of Excellence in Physical Chemistry, University of Peshawar, Peshawar 25120, Pakistan
b
Department of Physics, University of Agriculture, Faisalabad 38040, Pakistan
c
Institute of Horticultural Sciences, University of Agriculture, Faisalabad 38040, Pakistan
d
Laser Spectroscopy Lab, Department of Physics, University of Agriculture, Faisalabad 38040, Pakistan

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

Article history: Effect of pre-sowing magnetic field (MF) treatments on germination, seedling growth and yield attributes
Received 3 September 2015 in bitter gourd (Momordica charantia L.; cv Faisalabad Long) was investigated. Genetically uniform seeds
Received in revised form were exposed to 25, 50, and 75 mT generated by an electromagnet (rectified sinusoidal non-uniform) for
10 November 2015
15, 30 and 45 min each. The treated seeds were sown and grown under field conditions in experimental
Accepted 3 December 2015
Available online 6 December 2015
plots (2.4 m2) according to standard agricultural practices along with control. The germination, growth,
and yield parameters were measured by standard methods and for mineral contents, laser induced
Keywords: breakdown spectroscopy (LIBS) was used. The enhancements in germination, emergence index, mean
Magnetic field germination time (MGT) and vigor index I and II were recorded up to 54.52%, 50.92%, 35.98%, 24.93% and
Pre-sowing seed treatment
47.21%, respectively. The growth parameters i.e., growth rate (56.98%), leaf area (64.61%), root fresh
Bitter gourd
weight (55.77%), root dry weight (76.16%), shoot dry weight (45.26%), shoot fresh weight (23.35%), shoot
Seedling growth
Yield length (29.39%) and root length (17.78%) were also enhanced significantly in plants, raised from mag-
Laser induced breakdown spectroscopy netically treated seeds. The chlorophyll contents (35.41%), fruit length (18.11%), fruit weight (14.93%),
yield (29.16%) and mineral contents were also recorded to be higher in MF treated plants group versus
control. Results suggest that pre-sowing MF treatment could possibly be used to improve the pro-
ductivity by enhancing germination and seedling growth since this treatment is eco-friendly, affordable
and applicable at cultivator level.
& 2015 Elsevier Ltd. All rights reserved.

1. Introduction
Bitter gourd (Momordica charantia L.) or Karela (Pakistan),
which belongs to family Cucurbitaceae, is used as a food and also
has medicinal importance. Bitter gourd grows in various tropical
regions of the world i.e., India, Malaya, China, tropical Africa,
Middle East, America (Kubola and Siriamornpun, 2008) and it is
one of the most important summer vegetables in Pakistan as well.
Generally, bitter gourd growing season in Pakistan and India is
January to June and now-a-days, it is usually grown as an annual
crop (Saleem et al., 2014). Optimum temperature of 25
28 °C is
required for the germination of bitter gourd seeds and in Pakistani
region temperature remains high except short winter season. The
mildew that often occurs during germination is responsible for
uneven and low germination rates (Peter et al., 1998). Biological
n
Corresponding author.
E-mail address: bosalvee@yahoo.com (M. Iqbal).
and chemical pre-sowing seed treatments have been used for
better seed germination and these treatments are considered cost
effective and also harmful to the environment (Iqbal et al., 2012a)
and the environmental safety is also an important issue world-
wide, which already has been contaminated and polluted with
toxic agents due to urbanization and industrialization (Abbas et al.,
2015; Adesola et al., 2016; Babarinde and Onyiaocha, 2016; Bilal
et al., 2014; Iqbal et al., 2013, 2014; Iqbal, 2016; Iqbal and Bhatti,
2014, 2015; Iqbal and Khera, 2015; Iqbal and Nisar, 2015; Jamal
et al., 2015; Manzoor et al., 2013; Qureshi et al., 2015; Sayed, 2015;
Ukpaka, 2016; Ukpaka et al., 2015; Ullah et al., 2013; Younas et al.,
2015). Therefore, it is important to adopt sustainable agricultural
practices i.e., environmental friendly, affordable and safe methods
(Iqbal et al., 2013; Majkowska-Gadomska et al., 2015; Maqsood
et al., 2013).
Modern agricultural efforts are now in search of a competent
ecological tool based on physical treatment of seeds to enhance
the germination, seedling strength and crop yield and recently,

http://dx.doi.org/10.1016/j.bcab.2015.12.002
1878-8181/& 2015 Elsevier Ltd. All rights reserved.
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37
pre-sowing MF seed treatment has attracted the attention of
agricultural community (Iqbal et al., 2012a; Jamil et al., 2012;
Kowalyszyn and Konyk, 2015; Naz et al., 2012; Zia ul Haq et al.,
2012). Various researchers have studied and reported that vege-
table and grain crops seed treated by magnetic field showed high
performance in terms of germination, plant growth, height, seed
mass per spike as well as shoot and root length and total fresh and
dry masses, chlorophyll contents, enzymatic activities and yield
(Iqbal et al., 2013; Iqbal et al., 2012a; Jamil et al., 2012; Jamil et al.,
2013; Naz et al., 2012; Perveen et al., 2011; Zia ul Haq et al., 2012).
There are numerous hypotheses explaining cellular responses of
MF on biological systems; however, precise mechanism is not well
known. Based on research conducted in this field, particles having
ferromagnetic properties in living organism, energy level amend-
ment and changes in electron spins in atom and molecules are
considered important. Different researchers presume that MF can
effect chemical reactions by altering electron spin location and in
this way they have potential to cause biological effects (De Souza
et al., 2014; Iqbal et al., 2012a). Electromagnet induced physiolo-
gical and biochemical changes in biological objects. Water assim-
ilation and intensified photosynthesis collectively enhance the
seed germination and growth (Podleoeny et al., 2004). In view of
importance of bitter gourd, nevertheless attention has been paid
to evaluate the effect of MF of germination, growth and yield since
it is a common vegetable in Pakistan. Therefore, the principal
objective of the present study was to appraise the MF pre-sowing
seed treatment effect on germination, seedling growth, mineral
contents (by LIBS) and yield characteristics of bitter gourd native
to Pakistani soil.
2. Material and methods
2.1. Soil description and experimental area
The experiment was conducted in March, 2013 at the Post-
graduate, Agriculture Research Station, University of Agriculture,
Faisalabad (31°.26′ N, 73°.06′ E). The soil was of sandy clay loam in
texture, pH 8.1, pH water 6.67, organic matter (0.87%) and elec-
trical conductivity (2.6 dSm
1). The meteorological data regarding
rainfall, relative humidity and temperature were recorded from
meteorological observatory cell in the immediate vicinity of the
field during the experimental period. The average temperature,
relative humidity, PAN evaporation, sun shine and wind speed
values were recorded to be 20 °C, 59.9%, 03.5 mm, 8.4 h and
5.8 km/h, respectively (March, 2013), whereas these values were
25.1 °C, 47.0%, 05.9 mm, 9.3 h, and 7.2 km/h, respectively (April,
2013).
2.2. Seed collection and MF treatment
The bitter gourd (Momordica charantia L, cv Faisalabad Long)
authentic seeds were obtained from Ayyub Agriculture Research
Institute, Faisalabad, Pakistan. Uniform seeds were selected by
hand picking for MF treatment. Seeds having visible defects, insect
damage or malformation were discarded and selected seeds were
stored in desiccators over 70% (v/v) glycerin. The procedure and
electromagnet specification has already been reported in detail
elsewhere (Iqbal et al., 2013; Naz et al., 2012). Total nine different
MF doses (strength and exposure time) were selected for seed
treatment i.e., T1 ¼25 mT for 15 min, T2 ¼25 mT for 30 min, T3
¼25 mT for 45 min, T4 ¼50 mT for 15 min, T5 ¼50 mT for 30 min,
T6 ¼50 mT for 45 min, T7 ¼75 mT for 15 min, T8 ¼ 75 mT for
30 min, T9 ¼ 75 mT for 45 min and T0 ¼ Control (untreated seeds).
31
2.3. Seed sowing and culture practices
The experiments were carried out in Randomized Complete
Block Design (RCBD) in triplicate. The plot size was kept 1.5 m x
2 m (4.5 m2). Row to row and plant to plant distance was 1.5 m
and 60 cm, respectively. Seeds were seeded in pit at 2.5–3.0 cm
depth. First irrigation was applied after 10 days and then, regularly
on weekly basis in such a way that the moisture contents
remained 480%. Three hoeing were given to keep plots free from
weeds during the early stage of growth. All treatments in the ex-
periments were run simultaneously along with control under si-
milar conditions.
2.4. Germination measurement
For germination, ISTA (ISTA, 2004) guidelines were adopted.
The germination percentage of bitter gourd was measured fol-
lowing the reported method (Carbonell et al., 2000) and percen-
tage germination was calculated using the relation shown in Eq. 1.
Seed germinated
Germination (%) = × 100
total seeds (1)
Emergence rate index (ERI) was calculated as described in the
association of official seed analysts (Grewal and Maheshwari,
2011) using the relation shown in Eq. 2, where X1, X2, …Xn are the
number of seedling emerged on 1st, 2nd,… nth day after sowing, N
is the days taken for germination and np is presenting germinated
seed.
⎧X +X +X…+X ⎫
ERI (%) = ∑ ⎨ 1 2 3 n
⎬
⎩ N (np ) ⎭ (2)
Mean germination time (MGT) was measured using the relation
shown in Eq. 3, where n is the number of seeds germinated on day
D, D is the number of days counted from the beginning of ger-
mination and Ʃn is the final germinated seed. Vigor indices I and II
of the seeds were calculated by relations (Eqs. 4–5) (Vashisth and
Nagarajan, 2010).
∑ Dn
MGT =
∑n (3)
Vigor Index I = Germination (%) × seedling length (root +shoot) (4)
Vigor Index II = Germination (%) × seedling dry weight (root +shoot) (5)
2.5. Growth parameter measurements
The growth rate was calculated using the relation shown in Eq.
6. Leaf area was measured by using portable leaf area measure-
ment meter (Model YMJ-A, China) and averages were computed
by statistical analysis. The root lengths (cm), root fresh and dry
weight (g), shoot fresh and dry weight (g) were calculated after 45
days of sowing. Dry weights (root and shoot) were determined
after heating at 105 °C for 15 min, followed by incubation at 80 °C
for 24 h. The root length was determined by scale from root neck
to tip (Iqbal et al., 2013; Iqbal et al., 2012a; Iqbal et al., 2012b; Zia
ul Haq et al., 2012). All measurements were performed in triplicate
and data thus, obtained was averaged.
Plant height at maturity
Growth rate (%) =
Number of days taken (6)
32 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37
2.6. Chlorophyll contents
Fresh leaves, 45-day-old (0.1 g) were grinded in 1 mL acetone
and volume was made to 5 mL by adding more acetone. Then, after
being centrifuged for 5 min, supernatant was collected and the
absorbance was measured at 645 and 663 nm (CE Cecil 7200, UK).
The chlorophyll “a” (chl a), chlorophyll “b” (chl b) and total
chlorophyll (total chl) contents were measured using relations
(Eqs. (7)–(9)), respectively, where, V is the volume, W is the weight
of fresh leaves taken and A is the absorbance (Maqsood et al.,
2013).
⎧ 12. 7 (A663) –2. 69 (A645) ⎫
Chl a = ⎨ ⎬×V
⎩ 1000 × W ⎭ (7)
⎧ 22. 9 (A645) –4. 68 (A663) ⎫
Chl b = ⎨ ⎬×V
⎩ 1000 × W ⎭ (8)
⎧ 20. 2 (A645) + 8. 02 (A663) ⎫
Chl total = ⎨ ⎬×V
⎩ 1000 × W ⎭ (9)
2.7. Yield and mineral estimation
Fruit length and weight was measured by randomly selecting
mature nine fruits from each treatment by scale and electronic
balance, respectively and both were averaged. For total yield,
mature fruits from sub-plots were collected and weighed reg-
ularly, averaged and converted into yield/acre (kg). The mineral
contents were estimated by LIBS technique. For this, dried plants
were grinded and the mineral contents were estimated using laser
induced breakdown spectroscopy technique (Pouzar et al., 2009).
Pulsed Q-switched Nd:YAG (Quantel Brilliant) laser setup was
used. The laser setup for this experimentation is shown in Fig. 1.
The Nd:YAG laser with an ability of delivering 200 mJ at 1064 nm
wavelength was used. The emission spectrum was recorded with
the help of spectrometer. In order to record the emission spectrum
from the plume generated, the LIBS detection system and Q-switch
of the Nd:YAG laser was synchronized (Shaikh et al., 2006).
2.8. Statistical analysis
All treatments were performed in triplicate under RCBD and
data regarding germination, growth and yield characteristics were
statistically analyzed by one-way ANOVA (Po 0.05) at 95% con-
fidence level.
Fig. 1. Laser breakdown spectroscopy setup.
3. Results and discussions
Bitter gourd (Mamordica charantia L.) is one of the most im-
portant vegetables in Pakistan. It has been reported that bitter
gourd has low germination and field emergence due to thick coat
and unfavorable environmental condition i.e., mildew during
germination and variation in temperature (Peter et al., 1998;
Saleem et al., 2014). Present research was conducted to investigate
the effect of MF pre-sowing seed treatment on the germination,
growth, and yield attributes of bitter gourd. The trials were con-
ducted from March, 2013 to onward for the period of 100 days on
Faisalabad Long cultivar of bitter gourd, commonly grown in Pa-
kistan and results thus obtained are shown in Tables 1 (germina-
tion and growth parameters) and Table 2 (chlorophyll contents
and yield), Table 3 (analysis of variance-ANOVA) and Table 4
(percentage effect).
Germination response of MF treated seed revealed that MF pre-
sowing seed treatment enhanced germination characteristics. The
percentage germination of bitter gourd seed in T2, T4, T5, T7, T8 and
T9 effects was highly significant (P o0.05), whereas T1, T3 and T6
effect was statistically insignificant (P 40.05) and slightly higher
than control. The germination percentages were recorded to be
29.3%, 27.76%, 26.36%, 31.91%, 29.31%, 36.06% and 16.4% for T2, T3,
T5, T7, T8, T9, and T0, respectively and T8 enhanced the germination
percentage up to 54.52% higher than control (un-treated seed).
Previous studies also revealed that the MF treatment is helpful in
enhancing the germination and similar trend was observed in
present investigation e.g. the speed of germination of cucumber
seeds has been reported higher in seed treated at various MF doses
(Anand et al., 2012). Similarly, Flórez et al. (2007) treated maize
seed to the MF doses of 125 mT and 250 mT and resultantly, faster
germination and early growth was observed versus non-treated
seeds.
ERI indicated the rate of emergence of the seedlings and it was
enhanced in response of MF treatment. It was observed that T9, T8,
T7, T5, T4, T3 and T2 showed higher ERI values (P o0.05), whereas
T1 and T6 effect was insignificant (P 40.05) in comparison to
control. The maximum increment of 50.92% in ERI was observed
for T9 MF dose. The Coefficient of variance (CV) for ERI was 28.01,
which might be due to the lack of uniform experimental condi-
tions since experiments were conducted under field conditions.
These findings are consistent with previous studies (Grewal and
Maheshwari, 2011; Podleoeny et al., 2004). Authors reported sig-
nificantly higher ERI in broad bean seeds exposed to MF.
Regarding MGT, T3, T5 and T8 showed higher values, whereas T1,
T4, T5, T6, T7 and T9 showed low MGT and statistical significance
(P o0.05) with 32.78 CV values, which highlighted that MF treat-
ment might reduce the time taken for germination. The T5 treat-
ment reduced the MGT of bitter gourd seeds up to 35.98%, which
indicates that magnetically treated seed took 35.98% less time to
germinate as compared to untreated seeds. The effect of MF on
seed germination of two wheat cultivars has been studied (Gho-
lami et al., 2010) and MGT in both cultivars significantly decreased
when seed was treated for 2–3 h. In another study, effects of the
MF effect on maize seeds are reported and it was observed that
germination and growth was higher along with lower MGT value
versus control (Flórez et al., 2007).
The vigor index I values in T9, T8, T7, T4, and T2 treated groups
were 1873, 1721, 1600, 1570 and 1752, respectively which were
significantly (Po 0.05) higher than control, whereas in case of
control, the vigor index I value was 1406 and T1, T3, T5 and T6
showed the vigor index I values comparable with control. The CV
value of vigor index I was 44.49 and maximum enhancement in
vigor index I of 24.93% was recorded for T9. These results are
comparable with previous study (Vashisth and Nagarajan, 2010).
Authors studied the effects of static MF on the germination and
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37 33

Table 1
Effect of magnetic field treatment on germination and seedling growth parameters of bitter gourd.

Parameter Unit T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 CV

Germination % 16.4 19.6 29.3 20.5 27.76 26.36 18.64 31.91 29.31 36.06 27.4
7SD 0.60 0.80 1.00 0.85 1.10 1.15 0.40 1.40 1.60 1.00
Growth rate % 0.4 0.71 0.9 0.93 0.89 0.89 0.72 0.71 0.81 0.71 28.01
7SD 0.02 0.04 0.05 0.06 0.04 0.05 0.04 0.03 0.04 0.02
ERI 0.008 – 0.013 0.01 0.0123 0.012 0.0077 0.0143 0.0133 0.0163 37.42
7SD 0.001 – 0.01 0.01 0.001 0.01 0.000 0.01 0.01 0.01
MGT day 4.95 3.87 5.55 6.41 4.14 3.64 4.69 4.99 5.51 4.66 32.78
7SD 0.40 0.20 0.20 0.30 0.15 0.20 0.25 0.30 0.40 0.20
VI (I) 1406 1280 1752 1162 1570 1484 1335 1600 1721 1873 44.49
7SD 75 50 90 60 80 60 40 95 110 30
VI (II) 362 973 – 1404 1222 820 563 1137 1676 877 48.88
7SD 20 60 – 40 60 55 70 50 44 28
2
Leaf area cm 133.3 295.8 239 209.8 290.4 234.5 200.3 376.7 264.5 261.4 35
7SD 10 17 09 12 20 10 08 25 11 18
Root length cm 16.69 20.3 17.64 17.64 17.73 18.11 16.66 16.47 18.44 19.47 26.01
7SD 0.8 0.7 0.6 0.77 0.65 0.44 0.8 0.90 0.66 0.66
Shoot length cm 33.17 41.36 46.98 42.99 40.07 41.2 34.01 33.18 38.63 34.11 31.5
7SD 1.5 1.2 1.4 1.9 1.0 1.8 1.2 1.2 1.3 1.5
RFW g 4.63 10.47 8.1 10.47 8.87 7.33 6.33 9.43 – 5.77 24.89
7SD 0.4 0.3 0.4 0.45 0.6 0.4 1.0 0.9 – 0.1
SFW g 44.76 58.4 45.4 52.5 48.03 39.7 41.67 52.67 46.76 58.23 26.54
7SD 1.0 2.40 1.90 2.8 1.2 1.8 1.1 2.3 2.8 2.5
RDW g 0.472 1.45 1.23 1.51 1.13 1.33 1.04 1.7 1.98 0.95 26.06
7SD 0.04 0.08 0.06 0.08 0.09 0.05 0.04 0.10 0.15 0.09
SDW g 2.833 3.8 2.97 5.17 2.83 3.13 5.5 3.97 4.63 4.37 30
7SD 0.20 0.25 0.10 0.40 0.50 0.25 0.33 0.40 0.30 0.28

T1 ¼ 25 mT for 15 min, T2 ¼ 25 mT for 30 min, T3 ¼ 25 mT for 45 min, T4 ¼50 mT for 15 min, T5 ¼ 50 mT for 30 min, T6 ¼ 50 mT for 45 min, T7 ¼ 75 mT for 15 min, T8 ¼ 75 mT for
30 min, T9 ¼ 75 mT for 45 min, T0 ¼Control (un-treated seed), ERI¼ emergence rate index, MGT ¼mean germination time, VI I and II ¼ vigor index I and II, RFW¼ root fresh
waeight, SFW ¼shoot fresh weight, RDW ¼ root dry weight, SDW ¼ shoot dry weight, CV ¼coefficient of variation

growth of sunflower seeds and it was revealed that vigor index
increased as a result of MF treatment. In another study, the effect
of stationary MF was studied and the vigor index value of maize
seed increased (Flórez et al., 2007). Similarly, vigor index II was
also increased in MF treated group of bitter gourd plants and for T0
to T9, the vigor index II values were recorded to be 362, 973, 1550,
1404, 1222, 820, 563, 1137, 1676 and 877 with 48.88 CV value.
Percentage increase in vigor index II was 47.21% for T3. All other
MF treatments enhanced the vigor index II significantly (P o0.05)
versus control. In earlier study, the effects of static MF on sun-
flower seeds has been reported and it was found that the vigor
index II increased as a result of MF exposure (Vashisth and Na-
garajan, 2010) and in another study the vigor index II also in-
creased in magnetically treated seeds of maize (Anand et al., 2012).
Growth rate is considered one of the important parameters to
enhance the productivity and in present investigation, all MF doses
enhanced the growth rate significantly (P o0.05) versus control, as
it can be seen in Fig. 2, the plant growth was significantly different
after 20 days of germination and this difference was more clearer
at lateral stages of growth and development. Maximum increase in
growth rate was recorded to be 56.98% in T3 group in comparison
to control. These findings were comparable with earlier studies
that MF influenced the plant growth and development at lateral
stages (Iqbal et al., 2013; Iqbal et al., 2012b; Jamil et al., 2012).
Similarly, early growth has been reported in maize plant in re-
sponse of 125 mT MF and 250 mT doses as compared to control
(Flórez et al., 2007).
Leaf area of bitter gourd plant leaf raised from magnetically
treated seeds was also enhanced. All the MF treatment significantly
(Po0.05) affected the leaf area; however, treatment T7 effect was
more pronounced and 64.61% increment was recorded for this
treatment versus control. This trend is in line with previous studies
(De Souza et al., 2014; De Souza et al., 2008). Authors applied MF
dose of 100 mT for 10 min and 170 mT for 3 min and resultantly,
leaf area of tomato plant leaves enhanced considerably. Similarly,
160 mT for 15 and 20 mT treatment of onion also enhanced leaf
area. Regarding root length, T0, T1, T2, T3, T4, T5, T6, T7, T8 and T9
showed the root lengths of 16.69, 20.30, 17.643, 17.637, 17.730,

Table 2
Effect of magnetic field treatment on chlorophyll contents and yield characteristics of bitter gourd.

Parameter Unit T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 CV

n
Chl a mg/g 10 10.33 10.02 10.24 10.00 10.03 15.4 15.42 11.05 13.2 21.5
7SD 0.70 0.92 0.40 0.21 0.34 0.90 0.22 0.33 0.76 0.76
Chl b mg/g 3.12 5.46 5.46 5.41 3.08 4.27 5.5 7.36 5.65 5.98 25.6
7SD 0.20 0.30 0.40 0.30 0.20 0.10 0.44 0.33 0.44 0.26
Total Chl mg/g 14.3 16.6 15.9 14.3 13.53 13.91 20.35 22.14 – 19.01 22.26
7SD 1.80 1.50 1.95 1.86 1.80 1.20 1.90 1.70 – 1.15
Fruit length cm 16.05 16.16 16.68 19.33 16.20 16.88 16.41 16.60 19.60 16.66 21.73
7SD 1.90 1.97 1.40 1.20 1.55 1.90 1.90 1.00 1.50 0.95
Fruit weight g 60.59 66.11 69.5 70.5 70.09 61.5 71 67.33 71.23 68.4 29.75
7SD 5.64 2.97 3.17 3.41 4.21 5.69 3.26 2.04 1.21 4.80
Yield/acre Kg 1700 1750 2000 1960 1990 2130 2220 – 2400 2250 21.40
7SD 200 170 210 160 100 200 322 – 340 325

n
chl ¼chlorophyll and other explanations are same as given in Table 1.
34 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37

Table 3
Analysis of variance (ANOVA) of germination, seedling growth and yield
parameters.

Parameters SS MS F-Value P- Value Remarks

Germination (%) 2653.51 294.86 3.89 Po 0.05 Significant

Growth Rate 0.6709 0.07454 1.57 – Significant
ERI 2.42E–04 2.69E–05 1.46 – Significant
MGT 19.1787 2.13096 0.85 – Significant
Vigor Index (I) 3495372 388375 0.96 – Significant
Vigor Index (II) 4808766 534307 1.5 – Significant
Leaf Area 233526 25947.3 7.99 – Significant
Root length 51.334 5.7038 1.56 – Significant Fig. 2. Bitter gourd plant after 20 days of germination (T1 ¼ 25 mT for 15 min,
Shoot length 617.83 68.647 0.74 – Significant T2 ¼25 mT for 30 min, T3 ¼ 25 mT for 45 min, T4 ¼ 50 mT for 15 min, T5 ¼ 50 mT for
Root fresh weight 200.174 22.2515 2.49 – Significant 30 min, T6 ¼50 mT for 45 min, T7 ¼ 75 mT for 15 min, T8 ¼ 75 mT for 30 min, T9
Shoot fresh weight 3487.73 387.525 1.31 P4 0.05 Insignificant ¼75 mT for 45 min, T0 ¼control (un-treated seed)).
Root dry weight 4.8173 0.53526 1.35 Po 0.05 Significant
Shoot dry weight 48.2187 5.35763 4.66 – Significant In the present investigation, the MF treatments affected root fresh
Chl a 147.556 16.3951 4.04 – Significant weight positively (Po0.05), the treatments T1 and T3 effects were
Chl b 46.2954 5.14394 2.57 – Significant
Total chl 585.928 65.1031 8.27 – Significant
more pronounced and values were recorded to be 55.77% and
Fruit length 94.508 10.5009 0.84 – Significant 55.77% higher in comparison to control. However, all other treat-
Fruit weight 3808.9 423.21 1.06 – Significant ment effects were also considerably higher as compared to control.
Yield 180.00 20.000 5.05 – Significant Earlier, 120 mT for 10 min or 80 mT for 5 min MF dose enhanced the
SS ¼ sum of square, MS¼ mean square ERI¼ emergence rate index, MGT ¼ mean
fresh and dry shoot weight, shoot length, root length, fresh and dry
germination time, chl ¼chlorophyll root weight and leaf area in tomato plants (De Souza et al., 2005).
Regarding shoot fresh weight (g), T0, T1, T2, T3, T4, T5, T6, T7, T8 and T9
18.113, 16.657, 16.467, 18.443 and 19.47 (cm), respectively. Among values were found out to be 44.76, 58.4 (23.35%), 45.4 (1.44%), 52.5
treatments, T1, T5, T8 and T9 responses were significant (Po0.05) (14.74%), 48.03 (6.8%), 39.7 (
12.74%), 41.67 (
7.93%), 52.67
and others were insignificant as compared to control. The CV value (15.01%), 46.76 (4.27%) and 58.23 (g) (23.13% Vs T0). The CV value in
for root length was 26.01 and maximum increase in root length of case of shoot fresh weight was 26.54. Regarding root and shoot dry
17.78% was observed for T1. Similar to this study, sunflower root weights, all treatment furnished positive effect and resultantly, a
length increased as a result of MF treatments of 250 mT for 1–4 h considerable higher dry weights were recorded. Maximum incre-
(Vashisth and Nagarajan, 2009). In another study, the MF treatment ment in root and shoot dry weights were 76.16% and 48.54% in T8
affected positively the root length in response of 120 mT for 10 min and T6, respectively. Similar trend has been reported previously in
or 80 mT for 5 min treatments (De Souza et al., 2005). The shoot maize crop in response of 0.15 T MF treatment, which increased the
length of bitter gourd also recorded to be higher in plants raised shoot fresh weight up to 72% (Aladjadjiyan, 2002). In another study,
from magnetically treated seeds. The T1, T2, T3, T4, T5 and T8 effect tomato seeds treated by 120 mT for 10 min or 80 mT for 5 min MF
was significant (Po0.05), whereas T6, T7 and T8 treatments affect doses also showed significant increment in fresh and dry shoot
the shoot length marginally. The CV for shoot length was 24.89 and weights, shoot length, root length, fresh and dry root weight and
29.39% increase in shoot length was observed for T2. Earlier, MF leaf (De Souza et al., 2005). In another study, higher root and shoot
treated lentil seeds showed higher shoot lengths after 7th and 14th dry weights; root length, root volume and root surface area in
days of germination (Aladjadjiyan, 2010). Similarly, the MF treat- 1-month-old maize seedlings raised from MF treated seeds has
ments of 120 mT for 10 min or 80 mT for 5 min led to an increased been reported (Vashisth and Nagarajan, 2009).
shoot length, root length, fresh and dry root weight, fresh and dry In order to check the influence of pre-sowing MF treatment on
shoot weight and leaf area of tomato plant (De Souza et al., 2005). the chlorophyll content, chl a, chl b and total chl were measured

Table 4
Percentage effect (increasing/decreasing) of magnetic field pre-sowing seed treatment on germination, growth and yield characteristics of bitter gourd.

n
Parameters T1 T2 T3 T4 T5 T6 T7 T8 T9

Germination 44.02 20 40.92 37.78 12.01 48.6 44.04 54.52 40.14

Growth rate 43.66 55.55 56.98 55.05 55.05 44.44 43.66 50.61 43.66
ERI –5.26 38.46 20 33.33 33.33 -3.89 44.05 39.84 50.92
MGT – 10.74 24.33 –19.56 –35.98 -5.54 0.8 10.16 –11.77
VI I –9.94 19.74 –21.66 10.44 5.25 -5.31 12.25 18.3 24.93
VI II 62.79 – 74.21 70.37 55.85 35.7 68.16 78.4 58.72
Leaf area 54.93 44.22 36.46 54.09 43.15 33.44 64.61 49.6 49.05
Root length 17.78 5.38 5.38 5.86 7.84 -0.018 –1.33 9.49 14.27
Shoot length 19.8 29.39 22.84 17.21 19.49 2.46 0.03 14.13 2.75
RFW 55.77 42.83 55.77 47.8 36.83 26.85 50.9 – 19.75
SFW 23.35 1.44 14.74 6.8 -12.74 -7.93 15.01 4.27 23.13
RDW 67.44 61.62 68.74 58.23 64.51 54.61 72.23 76.16 50.31
SDW 26.31 4.71 45.26 0 9.58 48.54 40.28 38.87 35.24
Chl a 3.19 0.19 2.34 0 0.29 35.06 35.14 9.5 24.24
Chl b 42.85 42.85 42.32 –1.29 26.93 43.27 57.92 44.77 47.82
Total chl 13.85 10.06 0.00 –5.69 -2.8 29.72 35.41 – 24.77
Fruit length 0.68 3.77 16.96 0.92 4.91 2.19 3.31 18.11 3.66
Fruit weight 8.34 12.82 14.05 13.55 1.47 14.66 10.01 14.93 11.41
Yield/acre 2.85 15 13.26 14.57 20.18 23.42 – 29.16 24.44

n
Explanations as given in Table 1 and “–” indicates not determined.
 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37 35

and analyzed statistically and treatment effects were found sig-

nificant. It was observed that the chlorophyll contents increased as
the MF dose increased, but to a certain extent. The chl a content
was 10 mg/g in control, whereas it was 10.33 (3.19%), 10.02 (0.19%),
10.24 (2.34%), 10.00 (0.0%), 10.03 (0.29%), 15.40 (35.06%), 15.42
(35.14%), 15.05 (9.5%), 13.22 (mg/g) (24.24% Vs T0) in T1 to T9, re-
spectively. In case of chl b, the overall trend was same, T4 showed
negative response and T5 (26.93%) showed a slight lower chl b
contents as compared to T1, T2 and T3 and then it was increased in
T8 treatment. Total chl contents trend was similar to chl a, T4 effect
was negative (
5.69%) and T7 showed maximum total chlorophyll
content, which was 35.41% higher than T0. Present findings were
consistent with previous studies that MF has positive effect on
chlorophyll contents (Yao et al., 2005). Author reported effect of
MF on cucumber (Cucumis sativus) plant chlorophyll contents and
found that 0.2 and 0.45 T MF dose enhanced the cucumber leaf
chlorophyll content significantly. However, pea seeds exposed to
60, 120, 180 mT for 5, 10 and 15 min MF doses did not enhance the
chlorophyll contents (Iqbal et al., 2012a) and this difference might
be due to the crop and MF dose difference.
The MF pre-sowing seed treatment effect was also evaluated on
yield and yield components i.e., fruit length, fruit weight and
overall yield of bitter gourd. The average bitter gourd fruit lengths
were recorded to be 16.05 cm, 16.16 (0.68%), 16.68 cm (3.77%),
19.33 (16.96%), 16.20 cm (0.92%), 16.88 cm (4.91%), 16.41 cm
(2.19%), 16.60 cm (3.31%), 19.60 cm (18.11%) and 16.66 cm (3.31%
higher versus T0) in T0, T1, T2, T3, T4, T5, T6, T7, T8, T9 treatments,
respectively with 28.01 CV value for fruit length. Similarly, incre-
ment in fruit weight was also found significantly (Po 0.05) higher
in plants raised from magnetically treated seeds. In case of T1, T2,
T3, T4, T5, T6, T7, T8, T9 treatments, the fruit weights were recorded
to be 66.11 g (8.34%), 69.50 g (12.82%), 70.50 g (14.05%), 70.09 g
(13.55%), 61.50 g (1.47%), 71.00 g (14.66%), 67.33 g (10.01%), 71.23 g
(14.93%), 68.40 g (11.41% higher versus T0), respectively. The
maximum yield was recorded in T8 treatment, which was calcu-
lated to be 2400 kg/acre and all other treatments also yielded
significantly higher yield except T1 that was comparable with T0.
Overall, 2.85%, 15%, 13.26%, 14.57%, 20.18%, 23.42%, 29.16% and
24.44% higher yield was recorded in T1, T2, T3, T4, T5, T6, T8 and T9,
respectively. It has also been reported that the MF pre-sowing seed Fig. 3. Laser induced breakdown spectroscopy (LIBS) spectrum of bitter gourd
plant raised from magnetically treated seeds (A) T1 (25 mT for 15 min exposure)
treatment has accelerated effect on growth and yield of tomato
and (B) T9 (75 mT for 45 min exposure).
under field conditions. Author exposed tomato seeds to the MF
doses of 100 mT for 10 min and 170 mT for 3 min and resultantly,
fruit weight increased significantly (De Souza et al., 2006). In an- contents increased since intensity was higher in T9 as compared to
other study, the effect of pre-sowing MF treatments was in- T1. The studies regarding MF effect on mineral contents are rare;
vestigated on germination, growth and yield of okra for MF doses however, the effects of MF on mineral content were reported for
of 99 mT for 3 and 11 min. Resultantly, a significant higher ger- strawberry (Eşitken and Turan, 2004). Short day strawberry cv.
mination, number of flowers per plant, leaf area, plant height at Camarosa was treated with MF strengths of 0.096, 0.192 and
maturity, number of fruits per plant, pod mass per plant and 0.384 T and grown under greenhouse conditions and higher N, K,
number of seeds per plant were observed (Naz et al., 2012). Ca, Mg, Cu, Fe, Mn, Na and Zn was observed in 0.384 T treated
The qualitative mineral analysis was performed using LIBS plant, but phosphorous and sulfur contents reduced.
technique that is an atomic emission spectroscopy. Atoms are The MF pre-sowing seed treatment of bitter gourd was found
excited to high energy states and when they come back to the promising in enhancing the germination, growth, chlorophyll
ground state, energy is released. The released energy is well de- contents, mineral content and yield under field conditions. Results
fined for the specific excited atom, and this characteristic process showed that the bitter gourd germination and other moieties can
utilizes emission spectroscopy for the analytical method. LIBS be enhanced by applying an appropriate combination of MF dose
employs the laser pulse to atomize the sample and leads to atomic because it was observed that the germination, seedling growth,
emission. LIBS atomizes only the small portion of the sample by chlorophyll contents and yield attributes increased for specific MF
the focused laser pulse, which makes a tiny spark on the sample. doses (strength and exposure time) i.e., 25 mT for 30 and 45 min,
Because of the short-life of the spark emission, capturing the in- 50 mT for 30 and 45 min and 75 mT for 15 and 30 min were found
stant light is a major skill to collect sufficient intensity of the more effective in comparison to other treatments in present in-
emitting species (Nasim and Jamil, 2014). The Nd:YAG laser with vestigation. The MF action mechanisms on seed germination and
an ability of delivering 200 mJ at 1064 nm wavelength was used seedling growth is not well known, however several theories have
and the responses thus obtained are shown in Fig. 3(AB). The LIBS been proposed, including biochemical changes or altered enzyme
spectrum revealed the presence of Na, Ca, Mg and Fe. It can be activities. Seed germination stimulation might be attributed to a
observed from spectrum that by increasing the MF dose, mineral combined effect of biochemical, physiological, metabolic and
36 M. Iqbal et al. / Biocatalysis and Agricultural Biotechnology 5 (2016) 30–37
enhanced enzymatic activities. It is considered that that MF
treatment alters cell membrane permeability and enables to
transfer water and energy signals into the cell (Reina et al., 2001)
and resultantly, metabolic pathways may be enhanced (Iqbal et al.,
2012a, 2012b). The enzymes which are necessary for seed germi-
nation at particular stages of germination were found higher in
magnetically treated seed during seed germination (Vashisth and
Nagarajan, 2010). Another theory stated that MF affects the bio-
logical objects by non-conventional spins, free radicals, liquids
crystals or mobile electron charges and chemically these free ra-
dicals are very active and accelerate biochemical processes during
germination (Podleoeny et al., 2004). The increased water ab-
sorption has also been reported which might be responsible for
enhanced germination (Reina et al., 2001), especially in case of
bitter gourd, thick coat is one of the reasons for low germination.
The enhanced growth of bitter gourd at lateral stage of develop-
ment might be attributed to the enhanced antioxidant enzymes
activities (Vashisth and Nagarajan, 2010) because enzymes are
responsible for the degradation of food reserves stored in seed
during germination and protect the seedling from oxidative stress
during growth (Iqbal et al., 2012a). The enhanced leaf area in
seedlings raised from magnetically treated seeds may result in a
greater interception of light and might be responsible for higher
chlorophyll contents and photosynthetic rate. Hoff (Hoff, 1981)
found an increased photosynthetic rate and influx of water as a
result of magnetic treatments. It has been reported that MF
treatment accelerates the ions movement across plasma mem-
branes and amino acid (Stange et al., 2002) and these increased
ion content, biosynthesis of chlorophyll and carotenes are useful
for seedling nutrition (Dhawi and Al-Khayri, 2009). The increased
yield might be attributed to the favorable germination, higher
growth and enhanced photosynthetic rate because these para-
meters are the pre-requisite of flowering, fruit formation and ul-
timately, yield. As observed in present investigation, the fruit
length and fruit weights were significantly higher in plants raised
from magnetically treated seeds and this individual increase in
fruit is responsible for overall enhanced yield. The photosynthetic
rate was also observed higher in plants raised from magnetically
treated seeds and resultantly, yield may increased. Similar to
present study, 28–51% increase in yield in tomato and 12.5–14.5%
in wheat has been reported (Naz et al., 2012; Pietruszewski and
Kania, 2010).
4. Conclusions
Results revealed that MF accelerated the germination para-
meters i.e., germination percentage, emergence index, mean ger-
mination time and vigor index I and II. The growth parameters i.e.,
root length, root & shoot dry & fresh weights and leaf area also
increased significantly as a result of MF treatment. The chlorophyll
contents (chl a, chl b and total chl), fruit length, fruit weight and
ultimately, yield were also recorded higher in plant raised from
magnetically treated seeds. The MF doses (strength and exposure
time) were found to be variable for different parameters. Gen-
erally, 25 mT for 30 and 45 min, 50 mT for 30 and 45 min and
75 mT for 15 and 30 min treatment showed better performance.
Based on results, the use of pre-sowing MF seed treatment is
suggested to enhance germination, seedling growth and yield at-
tributes of bitter gourd.
Acknowledgments
We are thankful to authorities of the Ayyub Agriculture Re-
search Institute and Department of Agronomy for providing bitter
gourd seed and space for experimentation, respectively.
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