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 Vol 45, No. 4;Apr 2015

Effect of variable magnetic field on physiological quality

of broccoli (Brassica oleracea L.) seeds

Rico Martínez F.1, Hernández-Aguilar C.1, Domínguez-Pacheco A.1,

Paniagua Pardo G.1, Cruz-Orea A.2, Tomás-Velázquez S.2
1
Instituto Politécnico Nacional, SEPI-ESIME “Zacatenco”, Unidad Profesional “Adolfo
López Mateos”, Col. Lindavista, México D.F., C.P. 07738, México.
2
Departamento de Física, CINVESTAV – IPN, A.P. 14-740, México D.F., C.P. 07360,
México.
*Author correspondence: Tel 57296000 Ext 54807, E-mail: clauhaj@yahoo.com

Abstract
The effects of four alternating magnetic fields with four periods of exposition on
germination and early seedling growth of broccoli seeds were studied under
controlled conditions. The magnetic field treatments of 0.42 mT by 7.5 and 10 min,
obtained the best results, increasing significatively (P<0.01) dry weight 13.6% in both
cases, compared to control, while their effect on germination velocity index and fresh
weight also were favorable but without significant differences as compared to control
and other treatments. The treatment of 55 mT by 5 min, significantly decreased final
germination percentage (P<0.05), germination velocity index (P<0.01) and dry
weight (P<0.01) in 14.4, 25 and 11.3 %, respectively compared to control, while in
the fresh weight and average hypocotyl length, also produced negative effects
although no statistical differences. Meanwhile, the magnetic field treatments of 17
and 55 mT by 10 and 7.5 min respectively, completely inhibited the germination of
broccoli seeds. The results suggest that low intensity variable magnetic field
treatments (0.42 mT, 60 Hz) by 7.5 and 10 min could enhance physiological quality,
especially on dry weight of broccoli seed.

Keywords: broccoli, variable magnetic field, physiological quality, biostimulation.

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Introduction

Enough and good quality food production, is a need of global relevance (Rochalska et al.
2011), because it is projected a 40% population increase to 2050, being necessary to increase
agricultural production in 70%, otherwise, 4% of developing countries will suffer
malnutrition (FAO 2011). This has caused that science community is looking for methods to
increase agricultural production, finding that the germination, growth, productivity and
quality of crops are determined by environmental factors (Matwijczuk et al. 2012), as well as
by health, physical and physiological quality of seeds (Pietruszewski & Kania 2010; Mahajan
& Pandey 2011). The physiological quality improvement has been reported by different
authors using different methods (Aladjadjiyan 2007) such as chemical (fertilization)
(Aladjadjiyan 2012; Noha 2011), foliar fertilization (Zepeda et al. 2002), physical stimulation
(Pietruszewski & Kania 2010), laser light (Hernandez et al. 2006;2008;2010;2011; Ćwinta &
Dziwulska-Hunek 2013), high intensity LED (Paniagua et al. 2013), magnetic fields
(Hernandez et al. 2007; 2009; Dominguez et al. 2010; Rico et al. 2014a; 2014b; Dziwulska-
Hunek et al. 2009; Afzal et al. 2012; Marks & Szecówka 2010), among others. Chemical
methods have contributed the most to food production, but they have shown to be harmful
(Dominguez et al. 2010; Rochalska et al. 2011). Moreover, different authors say that the
physical method of biostimulation in presowing of seeds with magnetic field is friendly to the
environment and human health (Vasilevski 2003; Hernández et al. 2007; Aladjadjiyan 2010).
The magneto-reception in plants can be explained as the energy transference in matter
containing free radicals, which causes increased load, causing biostimulation (Hernández et
al. 2009; Aladjadjiyan 2010; Domínguez et al. 2010; Afzal et al. 2012); stimulating protein,
carbohydrates and water into them (De Souza et al. 2006), accelerating the processes of
germination and growth (Pietruszewski & Kania 2010). So, different plants react
physiologically and biochemically to different intensities and exposure times of
electromagnetic fields as shown by various investigations.
In pea seeds (Pisum sativum L.) stimulated with rectified magnetic field (RMF), it was found
significantly positive effects on the germination velocity index, final germination and vigor,
especially with 60 and 180mT by 5min (Iqbal et al. 2012a), on dry weight of roots and shoots
using 180mT to 50Hz by 10 and 15 min respectively, and positive trends in their amount of
chlorophyll (Iqbal et al. 2012), as well as its length and fresh weight of roots and shoots with
180mT by 5, 5 and 10 min, respectively, also increased the chlorophyll content (Iqbal et al.
2013).
Biostimulation with variable magnetic field (VMF) of 5mT to 16Hz by 2h can increase the
emergence speed and sugar content in low vigor seeds of sugar beet (Beta vulgaris L.) and
significantly improve their chlorophyll content (Rochalska et al. 2009). The pre-sowing
treatment of pepper seeds (Capsicum annuum L.) with the two poles of a permanent magnet
(PMF) of 62μT by 8, 12 and 24h, significantly increased the germination velocity and plant
total length, observing that the south pole has greater effects than the north one (Nimmi &
Madhu 2009).
In other plants positive effects also have been found. Two varieties of hybrid maize (Zea
mays L.) stimulated with 480mT of VMF to 60Hz by 3min, increased dry weight for San

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Jerome variety at 16%, while for the San Juan variety increased 21% with 9min (Zepeda et
al. 2010), on the other hand, to variety CL-11 x CL-12, stimulation with 100mT to 60Hz by
7.5min increased vigor and physiological quality (Hernandez et al. 2009), while, with 560mT
to 60Hz by 30min increased its emergency speed, setting percentage and dry weight of
seedlings (Dominguez et al. 2010). Considering all the above, the aim of this study was to
determine the effects of variable magnetic field on the physiological quality and optical
absorption spectra in leaves of seedlings of broccoli plants (Brassica oleracea L.).

The broccoli contributes to feeding as well as preventing and combating obesity (Lee et al.
2009; FAO 2006), and some cancers (Keum et al. 2009; Wang et al. 2010); so in countries in
process of development is important to increase their consumption..

Material and Methods

The experiment was conducted at IPN - ESIME Zacatenco University, Mexico DF. Itsco®
broccoli seed (Brassica oleracea L.) variety Waltham 29 were used, homogenized with a 1.2
± 0.25mm screen, it was sized with a Stariett®, 125MEB vernier. The average weight of 150
seeds was 4.49mg, measured with a Velab® electronic balance, VE-1000 model.

The research was conducted in the fall of 2013, with an average temperature and humidity of
20.48˚C and 31.71%, respectively. In this research seventeen treatments were evaluated with
four replications of 30 seeds including control, under an experimental design randomized
complete blocks.

After being embedded for 24 h, the seeds were treated with four magnetic fields of 0.42, 17,
24 and 55mT, and four exposure times each one 1, 5, 7.5 and 10min, as seen in Table 2.
Seeds were treated with arrange of coils, placed within the area of greatest concentration and
more homogeneous magnetic induction (B), measured with F.W.Bell® Gauss/Teslameter,
5070 model.

Then, seeds were sown in 9cm diameter Petri dishes with 0.4mm thick filter paper,
previously moistened with 3ml of purified water, the dishes were distributed into two foil
coated shelves, lighted with four modify LED emergency lamps Sanelec®, SE-2165-90L
model, providing photoperiods of 12h, with an intensity of 700 lux per shelf, measured with a
Steren® light meter HER-410 model.

The test lasted 15 days. Treatments were irrigated with 1 ml of purified water the days 3, 6, 8,
11 and 13 after sowing. Daily germination (G) was evaluated, considering seeds with a
radicle length equal to or greater than 2mm. The last day of test, final germination (FG) was
determined considering normal seedlings with minimum length of 8mm, modified method of
ISTA (2011).

With data obtained, germination velocity index (GVI) (Hussein et al. 2011; 2012) and final
germination percentage (PGF) (Domínguez et al. 2010) were determined.

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After FG was determined, 5 normal seedlings from each treatment and replicate were
randomly taken, by grinding their leaves in 5 ml of ethyl alcohol; the mixture was placed in
amber bottles for subsequently obtaining the spectrum of optical absorption of each, with a
spectrophotometer UV -VIS (UNICAM ®, UV-300 model).

With the remaining normal seedlings, average hypocotyl length (AHL) (AOSA, 1983) and
fresh weight (FW) by repetition were determined using an electronic balance (VELAB®, VE-
1000). Finally, seedlings were dried into a RIOSSA® oven E-51 at 65°C for 72 h, getting its
dry weight (DW), with an OHAUS® bascule, Adventurer model.

Data obtained were subjected to a variance analysis using the GLM procedure of SAS (SAS
2002). Subsequently, it was applied the method of highly significant differences (Tukey) with
a lower level of error 5% (α<0.05).

Results and Discussion

According to the variance analysis shown in Table 1, statistically significant differences were
found in final germination percentage (FGP), germination velocity index (GVI) and dry
weight (DW), to which means comparison analysis was applied, finding the following
differences (Table 2).

In this investigation, it was observed that treatments MF2-T4 and MF4-T3 (17 and 55mT by
10 and 7.5min respectively), produced the greatest negative biostimulation, completely
inhibiting germination (Table 2 and Figure 1).

Table 1. Variance analysis to seeds treated with four magnetic fields and four exposure
times.
FV GL FGP (%) GVI FW (g) DW (g) AHL (mm)
Repetition 4 935.92** 166.26*** 0.103 ns 0.00055*** 89.79 ns
Lot 17 231.11 54.76 0.014 0.00018 41.39
Error 67 1296.66 144.81 0.27 0.0005 438.14
2
R 0.47 0.604 0.29 0.553 0.23
VC 6.34 9.24 13.96 8.407 12.11
Mean 87.55 20.07 0.583 0.044 26.66
FGP - final germination percentage, GVI - germination velocity index, FW - fresh weight,
DW - dry weight, AHL - average hypocotyl length. *** - highly significant (p<0.01), ** -
significant (p<0.05), ns - not significant (p> 0.05).

Regarding final germination percentage (PGF), it was observed that MF2-T2 and MF4-T2
(17 and 55mT by 5min) treatments produced negative biostimulation (P<0.05) reducing it by
11.7 and 14.4% respectively, compared to control (Table 2). Similarly, treatments MF3-T2,
MF2-T3, MF3-T4 and MF4-T2 (24, 17, 24 and 55mT by 5, 7.5, 10 and 5min respectively)
had negative effects, with highly significant differences (P<0.01) in germination velocity
index (GVI) decreasing them by 10.4, 11.8, 15.1 and 25% respectively, compared to the
control (Table 2). Behavior that can be seen in Figures 1 and 2, respectively.

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Table 2. Means comparison analysis to seeds treated with four magnetic fields and four
exposure times.
Treatment B(mT) T(min) FGP (%) GVI FW (g) DW (g) AHL (mm)
MF1-T1 0.42 1 84.167 ab 20.746 a 0.580 a 0.040 b 25.367 a
MF1-T2 0.42 5 85.833 ab 20.437 ab 0.595 a 0.044 ab 25.531 a
MF1-T3 0.42 7.5 90.000 ab 21.875 a 0.615 a 0.050 a 27.639 a
MF1-T4 0.42 10 95.833 a 22.076 a 0.660 a 0.050 a 26.857 a
MF2-T1 17 1 90.000 ab 21.219 a 0.557 a 0.043 ab 26.170 a
MF2-T2 17 5 81.667 b 21.224 a 0.545 a 0.042 ab 27.557 a
MF2-T3 17 7.5 83.333 ab 18.486 ab 0.602 a 0.042 ab 27.778 a
MF2-T4 17 10 0.000 c 0.000 c 0.000 c 0.000 c 0.000 c
MF3-T1 24 1 89.167 ab 21.331 a 0.640 a 0.045 ab 26.204 a
MF3-T2 24 5 85.833 ab 18.784 ab 0.542 a 0.046 ab 26.400 a
MF3-T3 24 7.5 87.500 ab 20.318 ab 0.555 a 0.043 ab 27.031 a
MF3-T4 24 10 87.500 ab 17.785 ab 0.605 a 0.045 ab 28.431 a
MF4-T1 55 1 86.667 ab 20.423 ab 0.590 a 0.044 ab 23.749 a
MF4-T2 55 5 79.167 b 15.716 b 0.485 a 0.039 b 25.817 a
MF4-T3 55 7.5 0.000 c 0.000 c 0.000 c 0.000 c 0.000 c
MF4-T4 55 10 89.167 ab 19.741 ab 0.605 a 0.048 ab 28.497 a
Control 0 0 92.500 ab 20.971 a 0.580 a 0.044 ab 26.979 a
Means with the same letter in a column are statistically equal (Tukey, 0.05). MF – magnetic
field, T - treatment time, B - magnetic field intensity, FGP - final germination percentage,
GVI - germination velocity index, FW - fresh weight, DW - dry weight, AHL - average
length of hypocotyl.

Figure 1. Effects of four magnetic fields and four times exposure on broccoli seeds
(Brassica oleracea L.) at final germination percentage compared to control.

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Figure 2. Effects of four magnetic fields and four times exposure on broccoli seeds
(Brassica oleracea) at germination velocity index compared to control.

Dry weight (DW) variable had highly significant statistical differences (P<0.01) with
treatments MF1-T3 and MF1-T4 (0.42mT by 7.5 and 10min respectively), increasing it
13.6% in both cases, compared to control, and a significant diminishing (P<0.01) with
MF1-T1 y MF4-T2 (0.42 and 55mT by 1 and 5 min respectively) of 9.09 and 11.3%
respectively, compared to control (Figure 3). While the average length of hypocotyl
(LMH) and fresh weight (FW) variables, showed no effect to magnetic fields.

Figure 3. Effects of four magnetic fields and four times exposure on broccoli seeds
(Brassica oleracea) at dry weight compared to control.

Figure 4 shows daily germination velocity trends of treatments that caused positive
(MF1-T3 and MF1-T4) and negative (MF4-T2) outcomes in dry weight, compared to
control.

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Figure 4. Daily germination velocity of the three most significant magnetic field
treatments on broccoli seed compared to control.

Figure 5 shows the optical absorption spectra from leaves of seedlings whose seeds were
treated in pre-sowing with magnetic field of 0.42mT at four exposure time applied. It is
possible to observe the spectra in the range from 310 to 710nm, where the treatment that
presented a positive trend was 1min exposure. While treatment by 7.5min showed a negative
trend when compared with the control sample spectrum.

Figure 5. Optical absorption spectra of broccoli plants pre-sow treated with

variable magnetic field of 0.42mT by 1, 5, 7.5 y 10min, compared to control.

In the present investigation the indicator variables of physiological quality of seeds

of broccoli were evaluated: final germination percentage, germination velocity
index, fresh weight, dry weight and average hypocotyl length and the optical
absorption spectrum of their seedlings, finding that the physiological quality for
such dicot seed, can be enhanced or inhibited by applying variable magnetic field,
according to the intensity and exposure time used.
The treatments with 0.42mT by 7.5 and 10min, significantly increased dry weight of
seedlings of broccoli 0.45g, as happened in pea seeds (Pisum sativum L.) stimulated
with rectified magnetic field of 180mT by 15min, with a significant increase of

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0.18g, compared to control (Iqbal et al. 2012). In GVI and FW variables, there were
no statistically significant differences, but showed positive trends with respect to
control. Meanwhile, FGP and AHL variables had positive trends but not significant
with times of 10 and 7.5min respectively.
Other authors have reported negative biostimulation with variable magnetic field, in
this research it was found that the treatment with 55mT by 5min, significantly
decreased FGP, GVI and DW with respect to control. Similar effects were found with
hybrid maize (Zea Maya L.) CL-4 X CL-1, stimulated with 60mT by 7.5min,
decreasing seedling emergence 16.2%, however with 30min increased emergence
14% (Hernández et al. 2009); in pea seeds (Pisum sativum L.) stimulated with 180mT
by 15min, also significantly decreased germination velocity index (Iqbal et al. 2012a)
and in dodder seedling (Cuscuta L.) significantly decreased dry weight 10% with a
magnetic field stimulation of 88μT by 12 and 24h (Balouchi & Modarres 2009).
In this research, also could be seen a complete inhibition of germination with
treatments 17 y 55mT by 10 and 7.5min, respectively. Similar results were found in
wheat seeds (Triticum aestivum L.) treated with permanent magnets of 125 and
250mT, slowing the average germination time in three hours, when compared to
control (Gholami & Sharafi 2010), phenomenon that could be used to eliminate or
retard the undesired plants germination.
While AHL variable, did not presented effects to treatments and exposure times
applied. Unlike bean seeds (Phaseolus vulgaris L. cv. Gina) stimulated with 4 and
7mT by 7d, where a seedlings length increase was observed according the magnetic
field intensity was increased, suggesting that its effect is related to the field strength
(Cakmak et al. 2010).
The results found in this investigation show that the best response to biostimulation
was obtained with low level magnetic induction (0.42mT), other reports have
indicated a linear behavior in magnetic biostimulation as Iqbal et al. (2013) who
found that magnetic fields of low strength for longer exposure times, as well as,
magnetic fields of high strength for short exposure times are more effective than
other treatments.
The optical absorption spectrum of seedlings stimulated in pre-sowing with 0.42mT
by 1min showed a growth trend over the control (Figure 5), which might suggest that
the appropriate intensity and exposure time to VMF could produce significant
increases in chlorophyll content, improving photosynthetic quality of plants, because,
the amount of chlorophyll is related to the amount of nitrogen in plants and nutrient
uptake (Rochalska et al. 2009). The pea seeds (Pisum sativum L.) chlorophyll amount
stimulated with 60mT by 10 and 5 min, also showed a tendency of increase over the
control, although no significant differences (Iqbal et al. 2012), however, embedded
seeds for 2h before magnetic treatment with 60mt by 15min , caused a significant
increase in chlorophyll relative to control (Iqbal et al. 2013), whereas, soybean seeds

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(Glycine max L.) treated with permanent magnetic field with a variation from 2.9 to
4.6mT, it was found an increased and decreased its content with 2.2 and 19.8s,
respectively, relative to control (Atak et al. 2007).
The results of this investigation showed positive and negative effects on broccoli
seed´s physiological variables treated with varying magnetic fields; behaviors that
could be due to the affectation on its spin, charge of their mobile electrons or free
radicals of seed latter are chemically very active and react rapidly in the presence of
oxygen to produce changes in biochemical and physiological processes in the
germination of seeds (Iqbal et al. 2012a). The results found coincide with those
reported by Hernandez (2009), who suggests the existence of several effects of
stimulation produced by the varying magnetic field: zero effect, positive effect and
negative effect.
The electromagnetic field effects on germination and growth of plants depends on
several factors such as the field type, intensity and exposure time (Mahajan & Pandey
2014) as well as seeds material properties, because each seed variety reacts
differently to the same electromagnetic treatment, so, it is important to continue doing
research to determine appropriate times and intensities that cause an increase in the
physiological quality of broccoli, benefiting production and nutritional quality.

Conclusions

Positive and negative effects were found on variables of final germination percentage,
germination velocity index, fresh weight, dry weight and average hypocotyl length
broccoli seeds var. Waltham29 (Brassica oleracea L.), stimulated with variable
magnetic field, which were in function of the intensity and duration of the applied
magnetic field.

The best results were produced with treatments of 0.42mT by 7.5 and 10min on dry
weight, causing a significant increase (P<0.01) of 13.6% in both cases, while the
variables of germination velocity index and fresh weight also obtained favorable
results, although without significant differences compared to control.

Effects of negative biostimulation were found with the treatment of 55mT by 5min,
significantly decreasing final germination percentage (P<0.05), germination velocity
index (P<0.01) and dry weight (P<0.01) at 14.4, 25 and 11.3% respectively,
compared to control, while the variables fresh weight and average hypocotyl length, it
also were found negative results , but without statistically significant differences.

Treatments with variable magnetic field of 17 and 55mT by 10 and 7.5min

respectively, completely inhibit germination of seeds treated in pre-sowing.

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