F. Aghamir et al.

/ Eurasian J Soil Sci 2016, 5 (1) 39 - 46

Seed germination and seedling growth of bean (Phaseolus

vulgaris) as influenced by magnetized saline water
Fateme Aghamir a, Hosseinali Bahrami a,*, Mohammad Jafar Malakouti a,
Saeid Eshghi b, Foroud Sharifi c
a Tarbiat Modares University, Faculty of Agriculture, Department of Soil Science, Iran
b Shiraz University, Faculty of Agriculture, Department of Horticultural Science, Iran
c Iran Soil Conservation and Watershed Management Research Institute, Iran

Abstract
Magnetized water is considered eco-friendly physical presowing seed germination.The aim of
this study was to evaluate the effects of magnetized watertreatments on bean (Phaseolus
vulgaris) germination under saline conditions (0, 25, 50, 75, 100 and 120 mM NaCl). This
experiment was performed as factorial in a complete randomized design (CRD) with three
replications. The results revealed that the roots and shoots length, fresh and dry weight of shoots
and roots and roots to shoots ratio, chlorophyll content index, water uptake, tissue water
contentwere significantly affected by magnetized water.Irrigation with magnetized water
Article Info significantly increased the physiologic factors such as germination percentage and index, vigor
index and salt tolerance index, compared to untreated control seeds.Mean germination time and
parameters T1, T10, T25, T50and T90 (required time for germination of one to 90 percent of seeds)
Received : 13.04.2015 were reduced significantly in all magnetized water treated plants in comparison to control.The
Accepted : 18.08.2015 results also demonstrated that magnetized water was conducive to promote the growth of bean
seedlings under saline conditions.
Keywords: Chlorophyll content index, physiological parameters, salinity, vigor index
© 2016 Federation of Eurasian Soil Science Societies. All rights reserved

Introduction
Magnetic and electromagnetic treatments arebeingused in agriculture, as a noninvasive technique, to
improve the germination of seeds and increase crops and yields (Martinez et al., 2009). A safe method for
enhancing seed germination includes the reasonable use of chemical additives and substitution by
appropriate physical treatment. In recent years, there are growing concerns of eco-friendly physical
presowing seed treatments by newly developed methods such as magnetic field (MF) (Jamil et al., 2012). The
effects of different MFs varies depending on the age during exposure as well as the induction of the field
(magnetic field strength), period of exposure and environmental conditions such as temperature, humidity,
etc. (Pang and Dang, 2008).
In the beginning of 1980s, Fujio Shimazaki, a Japanese researcher, reveal that some physical andchemical
properties of water changed when it pass through MF; hecalled the passed water through MF, "magnetized
water" (MW) which lead to special functions (Cai et al., 2009); however,many researchers reported that MW
could promote germination and early growth (Qiuet al., 2011). Hilal and Hilal (2000) showed that
comparision between MF and MW in different treatment seeds. Magneticfieldtreatments had doubled
germination compared to MW ones on pepper seeds; however, tomato and cucumber seeds had responded
more to MW rather than MF. Soaking seeds is a crucial factor to enable MF effects because of the water’s role

*Corresponding author.
Tarbiat Modares University, Faculty of Agriculture, Department of Soil Science, Iran
Tel.: +9844580500 E-mail address: bahramih@modares.ac.ir
e-ISSN: 2147-4249 DOI: http://dx.doi.org/10.18393/ejss.2016.1.039-046
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as MF mediator.Both MW and MF treatments have succeeded to given the highest germination percentage
for cucumber cousing 86% increase over control (Hilal and Hilal, 2000). Magnetization of both seeds and
water has come to be the most effective treatment in seed germination as compared with control (Selim,
2008). Function of MW depends on induction, types (dynamicorstatic), length, andconfigurations of
homogeneous and heterogeneous of MF, numbers of repetitions of the process (flow revalations), water
flowpressure, material of pipeand composition of dissolved salts (Vashisth and Nagarajan, 2010). Dynamic
and static MW treatment improved rice seed germination and significant differences compared to control
were only obtained for the dynamic method (Carbonell et al., 2000).
Extensive research has revealed that the effects of MW depend on a wide range of factors including
specialenvironmental conditions such as temperature, humidity, and salinity (Gutzeit, 2001). Despite, MW
improved crop resistance in the alkali and salt conditions (Dandan and Yan, 2013) and the mechanism of its
effects highly depending on the concentration and composition of the dissolved salts (Ibrahim Mohamed and
Mohsen Ebead, 2013), there is a relatively little scientific literature on germination mechanism of MW in
saline condition. The present study was conducted to analyze the effects of MW, with six different salt
concentrations, on bean germination and seedling growth parameters.
Material and Methods
The magnetized water was produced byan electromagnetic field generator (Water Clear Environment
Technologies Company-SB 150) with signal cable is changed any where from 2000 to 24000 times a second
produces an oscillating and complex modulating frequency wave from the produces an inaudible sonic
impluse magnetic field strength (0.05-0.5mT).
Seed germination tests were conducted as a factorial experiment arranged in a complete randomized design
(CRD) with three replications. Seeds were sterilized by sodium hypochlorite and distilled water, placed in
petri dishes on a wet filter paper. Each treatment category (100 seeds) was irrigated with 20 ml of MW and
non MW (0, 25, 50, 75, 100 and 120mM NaCl). Seeds germinated in an incubator at 25°C and a relative
humidity of 50% (ISTA, 2004). Water uptake percentage (WUP) was recorded for 15 hours calculated by the
formula 1 according to Mujeeb-ur-Rahman et al. (2008).
W 2 W 1 [1]
WUP  100
W1
Where W1= initial weight of seed, and W2= weight of seed after absorbing water in a particular time. The pot
experiment was conducted with the aforementioned treatments "MW and non MWin a cultivated media box
filled with peatmoss and perlite (1:1). For each treatment category, 100 seeds was sown at the depth of 2cm
with three replications. The boxes were placed in green house at 25°C. After 2 weeks, plants harvested and
then the growth paramerets, including the roots and shoots length (RL and SL), fresh and dry weight of
shoots and roots (FWR, DWR, FWS and DWS) and roots to shoots ratio (R/S) were measured with scale
0.001 g.
Dry weights were determined after samples were oven-dried at 70°C for 48 h. Chlorophyll content index
(CCI) of fresh seedling leaves was determined using a portable chlorophyll content meter (CCM-200, OPTI-
SCIENCES, Tyngsboro, MA,USA) based on absorbance measurements at 660 and 940 nm (Markwell et al.,
1995).
The tissue water content (TWC) was calculated based on equation 2 (Black and Pritchard, 2002).
Freshweight  Dry weight [2]
TWC  100
Freshweight
The physiological factors consisted of germination percentage (GP), germination index (GI), vigor index (VI),
salt tolerance index (STI), required time for germination of one to 90 percent of seeds (parameters T 1, T10,
T25, T50, and T90) and mean germination time (MGT) were calculated based on equations 3 and 4.
n [3]
GP  100
N
GI  
 ni T i  [4]
N
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Where n represents the number of newly germinating seeds, and N total number of seeds; n iis number of
seeds germinated at day Ti and Ti is number of days after starting the test (Panwar and Bhardwaj, 2005).
Seedling vigor index was calculated based on Abdul-Baki and Anderson (1973).
VI  I   GP %  Seedling Lenght  Root  Shoot  [5]
VI  II   GP %  Seedling Dry Weight  Root  Shoot  [6]
Seedling Dry Weight  Root  Shoot  atS x [7]
STI 
Seedling Dry Weight  Root  Shoot  atS 0

Where, Sorepresents control and Sx a given salt concentration.The rate of germination was assessed by
determining the required time for germination of 1, 10, 25, 50, 75 and 90% of seeds (T 1, T10 ,T25 ,T50, T90)
(Carbonell et al., 2008) and the mean time germinating (MTG) calculated according to the equation 8
(Panwar and Bhardwaj, 2005).
n [8]
MTG  ( i 100)
Ti

Where, Ti is the number of days after starting the experiment and ni represent the total number of
germinated seeds on the ith day.
Statistical Analysis
ANOVA test was used for analyzing the data by using Minitab software at 5% probality level.
Results and Discussion
Analysis of variance of bean seed germination parameters are presented in Tables 1 and 2, and Figures 1 and
2.
Table 1. Analysis of variance for growth and developmental parameters of corn under different salinity levels and
magnetized water regimes; RL and SL (roots and shoots length, FWR, DWR, FWS, DWS (fresh and dry weight of shoots
and roots), R/S (roots to shoots ratio), TWC (tissue water content), WUP (water uptake percentage, WT (water
treatment: magnetized and non-magnetized) and EC (electrical conductivity).
Source df RL SL FWR DWR FWS DWS
WT 1 273.903** 110.600** 8.107** 0.088** 6.003** 0.046**
EC 5 102.170** 42.357** 0.831** 0.008** 2.276** 0.015**
WT*EC 5 3.294 ** 6.736 ** 0.259** 0.003** 0.374** 0.002**
Source df FWP DWP Rf/Sf Rd/Sd TWC WUP
WT 1 28.062** 0.260** 0.968** 0.773** 1189.53** 785.401**
EC 5 5.850** 0.045** 0.006* 0.012* 431.22** 252.516**
WT*EC 5 1.029 ** 0.008 ** 0.173** 0.038** 12.12** 10.528**
** and *: significant at P=0.01, and P=0.05, respectively; df: degree of freedom.

Table 2. Analysis of variance for corn physiological parameters as effected by magnetized water regimes and salinity;
CCI (chlorophyll content index), GP (germination percentage), GI (germination index), VI (vigor index), STI (salt
tolerance index), MTG (mean time generation), T1,T10,T25,T50, T75,T90 (required time for germination of seeds from one
to 90%), WT (water treatment: magnetized and non-magnetized) and EC (electrical conductivity).
Source df CCI GP GI VI(I) VI(II) STI
WT 1 312.111** 1133.44** 6.996** 4489172** 1470.340** 1.046**
EC 5 97.019** 1178.24** 8.004** 2010007** 416.450** 0.332**
WT*EC 5 4.208 ** 27.78** 0.402 ** 144379 ** 57.490** 0.056**
Source df MTG T1 T10 T25 T50 T90-T10
WT 1 18.431** 20.026** 6.003** 20.25** 8.028** 4.694**
EC 5 1.454** 1.447** 2.306** 2.583** 0.628ns 10.094**
WT*EC 5 0.347 ** 0.187** 0.415 * 1.117 ** 0.494ns 0.228ns
**, * and, ns: significant at P=0.01, P=0.05 and no significant, respectively; df: degree of freedom.

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Growth parameters
Analysis of variance of the growth parameters including roots and shoots lenght, fresh and dry weight of
roots and shoots, roots to shoots ratio, tissue water content and water uptake percentage exposed to
magnetized water are shown in Table 1 and Figure 1.

Figure 1. Effects of magnetized saline water on growth parameters of bean germination

(M: Magnetized and N : Non magnetized water, S: Salinity)

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Roots and shoots length

Root length provides an important clue to the response of plants to salinity stress. Analysis of variance in
table 1 showed that shoot and root length of the magnetized saline water treated seedlings were higher than
seedlings treated by saline water. Salt stress inhibited the growth of root more than shoot in all treatments.
Root and shoot length decreased by increasing NaCl concentration. A marked reduction in RL and SL of non-
MW treatments was observed following salt stress (Figure 1a,b). The result showed that RL and SL of the
magnetized saline water treatments siginificantly were higher than control by 69 and 40%, respectively
(Table1). Mahmood and Usman (2014) reported similar results on maize seed germination, as in their
experiment, magnetized saline water significantly increased SL up to 10.24% compared to saline water.
Roots and shoots dry and fresh weight
Salt stress decreased FWR, DWR, FWS and DWS in non-MW treatments (Figure 1c,d,e,f). Reduction in weight
can be due to proportional increase in Na+ concentration which disturbs ionic and osmotic balances;
however, dry weights were not much affected compared to the fresh weights. A significant increase in FWR,
DWR, FWS and DWSoccured in the magnetized saline water treated seedlings as compared to the control
(Table 1). There were significant differences between treatments for roots to shoots ratio (R/S) (Table 1). As
NaCl concentration increased, it unequally affected roots and shoots weight. The result of R/S represented
that salt stress inhibited the growth of roots more than shoots in all non-MW treatments (Figure 1i,j). Other
authors have reported similar findings (Jamil et al., 2006). As NaCl levels increased the ratio of roots to
shoots dry weight remarkably decreased. Reduction of dry weights relatively depended on shoot or root
lengths. Along with increase in NaCl concentration in magnetized saline water treatments, the R/S gradually
increased (up to 75 mM NaCl) (Figure 1 i, j), then decreased; however, it was significantly higher than non-
MW treatments.
Tissue water content (TWC)
Magnetized water increased TWC significantly in compairison with non-MW treatments (Table 1 and Figure
1k). Along with increase in the salinity intensity, Kang and Saltvett (2002) reported that the osmotic
pressure leads to a reduction of water absorbance and inhibition cell division and differentiation;, which
adversely affects metabolic and physiological processes, and this causes more delay in initiation of
germination followed by prolonged seed germination duration, and ultimately reduces germination
parameter and seedling growth. It seems that MW decreases injurious effects of salinity by increaseing water
absorption.
Water uptake percentage
As NaCl concentration increased (up to 120 Mm) WUP decreased in all treatments (Figure 1l). Increase in
salinity level decreased germination parameters and seedling growth which are directly related to external
osmotic potential preventing water uptake. Bean seeds exposure to MW increased WUP significantly
compaired to non-MW treatments (Table 1). The result are in agreement with other researchers, which
indicates that MW treated seeds absorbed more water and absorbed it faster than untreated seeds (Kaya et
al., 2006); moreover, results of this study are in agreement with findings which lettuce seeds exposure to
static MF causes increase in water uptake rate (Reina et al., 2001). MF changes both osmotic pressure and
ionic concentrations in the membrane angles, which regulates the water absorbance (Azharonok et al.,
2009).
Physiological parameters
Analysis of variance of physiological factors included chlorophyll content index, germination percentage,
germination index, vigor index, salt tolerance index, mean germination time and required time for
germination of seeds from one to 90%, whichare shown in Table 2 and Figure 2.
Chlorophyll content index (CCI)
As NaCl levels increased, the CCI decreased remarkably (Figure 2a) due to enzymatic chlorophyll
degradation (Xu et al., 2000). Magnetized saline water significantly increases CCI as compared to non-MW
(Table 2). This result was in agreement with previous works as it has been reported that MW increased
chlorophyll content of wheat, flax, and lentil (Qados and Hozayn, 2010) byincreasing the activity of the
chlorophyll degrading enzyme: Chlorophyllase (Jamil et al., 2007).

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Figure 2. The bean physiological parameters as affected by magnetized water and salinity
(M: Magnetized and N : Non magnetized water, S: Salinity)

Germination Percentage(GP)
The analysis of variance showed magnetized saline water had significant effect on GP compared with non
magnetized saline water (Table 2). Previous studies have shown that static and dynamic MW improved GP of
rice seeds (Carbonell et al., 2000); moreover, it increased GP of pepper 35.8% over control (Ahamed et al.,
2013). Increasing the salinity level increases the time required for germination. Some studies have reported
that increasing the salt concentration delays the tomato seeds germination (Mohamed and Ebead, 2013).
Germination Index (GI)
Germination test showed magnetized saline water had a significant effect on GI in comparison to control
seeds (Table 2). The result confirmed the study of Mahmood and Usman (2014), who indicated
improvement of GI from 10.20 to 11.85 in seedlings treated with saline magnetized water in comparison
with the control. The enhanced speed of seedgermination does suggest that magnetized water hasacquired
some structural changes; perhaps promotingwater flow and bioactive molecules availability.
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Vigor index (VI)

Vigor Index (I) values based on seedling length and GP showed a trend similar to seedling shoots and roots
length in non-MW treated seedlings. In magnetized saline water treatments, as NaCl levels increased, the
VI(I) gradually decreased (Figure 2d); however, it was significantly higher than non-MW treatments. MW
improved VI(II) values based on seedling dry weight and GP (Table 2). Along with increase in NaCl levels,
VI(II) increased (up to 50 Mm NaCl) in all magnetized saline water treatments, then decreasded (Figures 1
and 2); however, it was significantly higher than non-MW treatments (Table 2).
Salt tolerance index (STI)
At different levels of NaCl, STI was significantly different and decreased with increasing in salinity intensity
(Figure 2f). The magnetized saline water had significant effect on increasing STI in comparison to control
seeds (Table 2). This results were in agreement with the findings of Dandan and Yan (2013) who indicated
that applications of magnetically saline water increased STI of wheat and guinea grass.
Mean time germination (MTG)
Magnetized saline water treatment had significantly lower MTG than the control treatments (Table 2 and
Figure 2g). Results are in agreement with the germination data obtained by Mahmood and Usman (2014)
that magnetized saline water reduced MTG from 9.15 to 8.73 and germination completed two to three days
earlier than control. Time parameters (T1 ,T10 ,T25 , T50, and T90) in magnetized saline water treatments
significantly were lower than control (Table2), indicated less time required for seed germination, and
germination speed is higher. Parameters T1 and T10 are related to the beginning of germination, this study
represents a progress in germination and a reduction in the induction phase in most of the magnetized
saline water treatments applied.
Conclusion
Results of the current study revealed the positive and significant impacts of magnetic water on physiologic
factors, and growth and development parameters of bean (Phaseolus vulgaris) comparison with the control.
The stimulatory effect of magnetic water on the growth in this research may be due to the increase in roots
and shoots growth. Therefore, as a simple and safe method, irrigation with magnetic water can be used to
improve plant growth and water use efficiency. The results also demonstrated that magnetized water
increased the bean seedling tolerance under salinity conditions. Generally, using magnetic water treatment
could be a promising technique for agricultural improvements but extensive research is required on
different crops.
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