ULTRASONIC STIMULATION OF THE DEVELOPMENT

OF LENTILS AND WHEAT SEEDLINGS

ANNA ALADJADJIYAN
Department of Mathematics and Physics, Agricultural University, 12, Mendeleev Str., 4000, Plovdiv,
Bulgaria, e-mail: anna@au-plovdiv.bg

Abstract. This paper presents an investigation of the effect of ultrasonic treatment on the
development of seeds from lentils (Lens culinaris, Med.) and wheat (Triticum aestivum). Ultrasonic
wave with frequency 42 kHz has been used for the treatment. Samples were exposed to ultrasound for
0, 1, 2, and 3 min. The germination energy (GE), germination (G), length of stems (SL) and roots
(RL), measured on the 7th and 14th day, as well as the total weight (TW) have been used in order to
evaluate the impact of ultrasound on plant development. The length of stems (SL) and roots (RL),
measured on the 7th and 14th day, as well as the total weight (TM) have shown dependence on the
exposure time, better expressed for the parameters measured on the 7th day. The effect of ultrasound is
stronger for lentils than wheat seeds.
Key words: Ultrasound, lentils, wheat, germination, stem, roots length, total weight.

INTRODUCTION

The use of physical methods for plant growth stimulation attracts more and
more the attention of agricultural producers as an alternative of chemical
amelioration, which offers ways to improve food quality without impairing its
safety.
Ultrasound is mechanical wave having frequency higher than 20 kHz. It is
established [13] that the treatment with ultrasound could change the state of the
substances and even accelerate the reactions. The chemical effects of ultrasound
are diverse and include substantial improvements in chemical reactions.
The application of ultrasonic stimulation for rising seed germination and
early stages plant development has been investigated for different cultures:
chickpea, wheat, pepper, and watermelon [6], corn [7], rice cells (Oryza sativa
Nipponbare) [9], pepper, tomatoes and cucumbers [10], fodder beans [11], radish
[12], carrot [1], ornamental trees [2, 3]. Ultrasonic treatment of seeds was used also
for industrial purposes like oil extraction and malts preparation [8, 14, 15, 16],
________________________
Received: May 2011;
in final form July 2011.
ROMANIAN J. BIOPHYS., Vol. 21, No. 3, P. 179187, BUCHAREST, 2011

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Anna Aladjadjiyan

since cell destruction under the shock of the ultrasonic wave with high intensity
facilitates extraction.
The application of the ultrasonic treatment to different seeds has resulted in
the establishment of the proper regime for increasing plant development and yield.
Obtained results have indicated that the effects of the treatment on seed
germination depend on frequency of ultrasonic wave and exposure time as well as
on the species and cultivars. Most of the authors recommended the treatment with
ultrasound of frequencies 15 100 kHz and exposure from 1 to 60 min.
In our previous works the effect of ultrasonic treatment with frequency 22
kHz and a power of 150 W on the germinating energy and germination of carrot
seeds (Daucus carota L.), cv. Nantes was studied [1]. The maximum effect was
established for 5 min treatment. Similar results have been obtained for some
ornamental species [2, 3].
The aim of the present work is to investigate the effect of treatment with
ultrasound with higher frequency on the development of lentils and wheat. The
intensity I of ultrasonic wave is related to its frequency [4]:
I=

2 A2
v
2

(1)

where is the density of the medium, A is the amplitude of the ultrasonic wave,
and v the velocity of the sound. Hence one can expect that ultrasonic wave with
higher frequency will have a stronger impact on the seeds. The energy E,
absorbed by the seeds, depends not only on the intensity I, but also on exposure
time t:

t =

E
I S

(2)

That is because in the present study shorter exposure times have been chosen
compared to those in the previous ones [1, 2, 3].
MATERIALS AND METHODS

Seeds of lentils (Lens culinaris, Med.,) and wheat (Triticum aestivum) have
been used for investigating the influence of ultrasonic treatment on the early stage
development of plants. Both plants have been chosen envisaging the opportunity to
explore possible relation of seed morphology with the effect of ultrasonic
treatment.
Ultrasonic treatment of seeds has been implemented with an apparatus
Carrera Sinus 2501 with ultrasonic frequency 42 kHz and power of 100 W.

Ultrasonic stimulation of the seedlings

181

Seeds have been distributed in four variants, respectively for lentils and
wheat. Each variant has been repeated in 10 replicates containing 10 seeds each.
Seeds in each variant have been exposed independently to ultrasound treatment for
different time: 0 min (control), 1 min, 2 min, and 3 min.
Seeds have been soaked in a plastic container with tap water and placed in
the marked center of the ultrasonic apparatus. After the treatment the seeds were
cultured in small plastic pots ( = 7 cm and h = 7 cm) on wet filter paper.
The experiments have been performed in March April 2011 under
laboratory conditions. The natural light cycle was 10 h light/14 h darkness with
daily temperature 21 2 C, night temperature 15 2 C.
In order to estimate the influence of the ultrasonic treatment on lentils and
wheat seeds next some criteria have been used:
The germination energy (GE) of seeds in %, determined on the 4th day after
the start of the experiment as a ratio of the number of germinated to the total
number of seeds for the corresponding variant;
germination (G) of seeds in %, determined on 7th (8th ) day as a ratio of the
number of germinated to the total number of seeds;
length of stems (SL) and main roots (RL) in mm determined on the 7th and
th
14 day;
total weight (TW) in mg determined on the 14th day.
Data were statistically processed using the method of Fisher of dispersion
analysis. The values reported in Tables 1 and 2 are mean values on 100
measurements for each parameter.
RESULTS AND DISCUSSION

The results of the experiments have been presented in Tables 1 and 2, and
illustrated with Figures 15. Results are presented as average value standard
error.
Table 1
Growth parameters of lentils seeds, treated with ultrasound at frequency 42 kHz
Exposure
time (min)
0 (control)
1
2
3

GE (%)

G (%)

8420
945

929
966

928
928

985
985

7th day
SL (mm)
RL (mm)
5013
15,62
61,99
269
937

1008

a p <0.001, b p <0.01, c p <0.05

364

25611
22315

14th day
RL (mm)
845
c
924,5
818

494

2246

896

SL (mm)
18921
b

TW (mg)
40214
42715
43914

41714

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Anna Aladjadjiyan

Table 2
Growth parameters of wheat seeds, treated with ultrasound at 42 kHz
Exposure
time (min)
0 (control)
1
2
3

GE (%)
982
1000
969
965

G (%)
906
946
866
925

7th day
SL (mm) RL (mm)
752
15,32
828
246
a
792
345
878

386

SL (mm)
2167
2067
2078
2096

14th day
RL (mm)
1867
b
1955
1856
1876

TW (mg)
37025
38226
37638
38321

a p <0.001, b p <0.01, c p <0.05

The germination energy (GE) and the germination (G) for seeds of lentils and
wheat are presented in Table 1 and Table 2, respectively. Germination energy show
the highest value for both lentils and wheat seed for exposure time 1 min. For these
samples (1 min US treatment) rise in GE is 11% for lentils and 2% for wheat.
Germination of lentils rises with exposure time for all the treated samples, while G
for wheat has its biggest value for exposure time 1 min. Values for G are lower for
wheat than for lentils. In the case of wheat the values for G are lower than those for
GE. Differences between variances and control are not statistically significant.

Fig. 1. Stem (SL) and root length (RL) of lentils seedlings exposed for different time
to ultrasound treatment, measured on the 7th day.

The results for the SL and RL for seeds of lentils measured on the 7th day are
illustrated in Fig.1. It can be seen that the values for treated samples both for
SL and RL rise with exposure time. The rise for exposure time 1 min is 24%,
for 2 min 86%, and for 3 min 100%.
In case of SL for wheat seeds (Fig. 2) the rises are not so expressive. It is
respectively 9% for 1 min, 5% for 2 min and 16% for 3 min exposure. The rise of
RL both for wheat and lentils seeds is well expressed. The values for lentils are
67% for 1 min exposure time, 130% for 2 min and 214% for 3 min. In the case of
wheat it is 56%, 122 and 148%, respectively.

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183

Fig. 2. Stem (SL) and root length (RL) of wheat seedlings exposed for different time
to ultrasound treatment, measured on the 7th day.

Fig. 3. Stem (SL) and root length (RL) of lentils seedlings exposed for different time
to ultrasound treatment, measured on the 14th day.

The results for the measurements on the 14th day show some differences
between the behaviour of lentils and wheat seeds. In the case of lentils SL (Fig. 3)
for treated samples are longer than the control one. The change is respectively 35%
for 1 min, 18% for 2 min and 18.5% for 3 min. A well expressed maximum is
accounted for exposure time 1 min.
In the case of wheat (Fig. 4) the longest SL is measured for untreated
(control) samples. The values of SL for treated samples are less than the control
one. The values of RL for samples treated at exposure time 1 min are bigger than
the control, both for lentils and wheat. The rises are 7% and 10% at exposure time
1 min, for lentils and wheat, respectively. In the case of wheat the difference is
statistically significant. For exposure time 3 min the rise 6% has been accounted
only for lentils.

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Anna Aladjadjiyan

Fig. 4. Stem (SL) and root length (RL) of wheat seedlings exposed for different time
to ultrasound treatment, measured on the 14th day.

The total weights of lentils and wheat seeds are presented in Fig. 5. From
Tables 1 and 2 it can be noticed that both for lentils and wheat the TW of all the
treated samples are bigger than control ones. The comparison shows that the TW of
the samples of lentils are bigger than those of wheat, but in the case of wheat the
difference between TW of treated and control samples is smaller than that for
lentils. For all the variants the differences are not statistically significant. The rise
of TW is 6%, 4%, and 9% for exposure times 1 min, 2 min, and 3 min, respectively,
in the case of lentils. In the case of wheat it is 3%, 2%, and 4%, respectively.

Fig. 5. Total weight (TW) of lentils and wheat seedlings exposed for different time
to ultrasound treatment, measured on the 14th day.

We have to point out that only the length of the main root has been measured
without taking into account the lateral roots. This can partially explain the
differences in the rise of TW for the samples, for which a rise of SL and RL was not
observed.

Ultrasonic stimulation of the seedlings

185

Because of the investigation of various seeds, with use of ultrasound sources

producing different ultrasonic frequencies and the variety of chosen times of
exposure described in the literature, it was difficult to compare the results obtained
in the present work with the previously published ones. No investigations on lentils
were found in studied literature. Results for wheat can be compared with those
announced by Goussous et al. [6]. In this study an ultrasound wave with frequency
40 kHz and output power 100 W has been used. The exposure times chosen for the
treatment were 5, 10, 15, 30, 45 and 60 min. The favourable effect on GE the
authors have found for exposure time 5 min. Most of the studies [14, 15, 16]
present treatment of investigated seeds with ultrasonic frequency 20 kHz compared
with 42 kHz in our investigation. Their results cannot be directly compared with
ours; nevertheless, they also confirm the stimulation effect of ultrasound on
germination of barley seeds.
Suslick [13] mentioned that in some cases, ultrasonic irradiation can increase
reactivities by nearly a million fold. The rise in plant growth characteristics for
both lentils and wheat, better expressed for the measurements on the 7th day, may
be explained with the increased reactivity of biological substances in the seeds.
Later measurements on the 14th day show that the differences between treated and
control samples decrease. We suppose that this fact may be explained with the
attenuation of the effect of ultrasonic treatment in time.
Another opinion is expressed by Liu et al. [9]. They have treated rice cells
with ultrasound with frequency 28 kHz for exposure time 2120 s and have found
the optimal stimulation of 5 s, while with longer treatment, the growth and
proliferation of rice cells has been inhibited. The authors suppose that the
ultrasound activates or destroys the cellular structure, such as cell membrane,
cytoskeleton and mitochondria in which many enzymes and ion channels are
affected. They assume that the enhancement of cell wall and cell membrane
fluidity might be one of the factors to promote the cell growth in 5 s ultrasonic
stimulation.
The weaker effect of ultrasonic treatment observed in TW of wheat seeds
compared with lentils may be due to a difference in seed coat thicknesses and its
physical properties. Fraczek et al. [5] investigated seed coat thickness of different
cultivars and differentiated two seed groups: one in which seed coat thickness
increases with increased moisture content, and another group in which seed coat
thickness decreases with increased moisture content (water permeable seed coat).
First group includes wheat, the second one includes beans. Lentil in morphological
aspect is similar to beans. Its seed coat decreases with increasing moisture content
and allows seed absorbing much water. Increased thickness of wheat seed coat
during its ultrasonic treatment is an obstacle for absorbing water and may cause
reducing the effect of ultrasonic stimulation. It could explain also the observation
that the values for G are lower for wheat than for lentils.

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Anna Aladjadjiyan

CONCLUSIONS

Results obtained in this investigation allow to conclude that ultrasonic

treatment with frequency 42 kHz of lentil and wheat seeds improves the early
stages of plants growth.
The growth characteristics SL and RL measured on the 7th day for lentil and
wheat increase quasi linearly with the increase of exposure time.
The best results on the 14th day have been achieved at exposure time 1 min
for the SL of lentil seeds and for RL of both lentil and wheat seeds.
The measured values of TW on the 14th day for treated samples both for
lentils and wheat are higher than the control ones. A linear dependence of TW vs.
exposure time may be accepted.
The weaker effect of ultrasonic treatment in TW of wheat compared with that
of lentil may be explained with the differences in physical properties of seed coat.
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