Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34

www.elsevier.com/locate/colsurfb

Biological effect of sound field stimulation on paddy rice seeds

Wang Bochu *, Chen Xin, Wang Zhen, Fu Qizhong, Zhou Hao, Ran Liang
Key Lab for Biomechanics and Tissue Engineering under the State Ministry of Education, Bioengineering College, Chongqing University,
Chongqing 400044, China

Received 19 March 2003; accepted 14 May 2003

Abstract

In this article, we adopt the apparatus invented by ourselves to investigate on the sound effects on paddy rice seeds in
the germination index, height of stem, relative increase rate of fresh weight, rooting ability, activity of root system and
the penetrability of cell membrane. The experiment results show that 400 Hz and 106 dB are the ‘best frequency and
intensity’. But when the sound wave stimulation is in excess of 4 kHz or 111 dB, it is harmful for paddy rice seeds. The
study not only validates sound field stimulation can certainly promote the growth of plant, but also indicates the
practicability of the apparatus.
# 2003 Elsevier B.V. All rights reserved.

Keywords: Sound field stimulation; Paddy rice seed; Biological effect

1. Introduction effect on many plants’ biological indexes [1
/7].

Living in the nature, plants are unavoidably
influenced by kinds of outside environmental
factors on quality and output, which are important
for human life and development. It has become an
inevitable task for scientists to study the effects
and action mechanism of the environmental fac-
tors.
In the growing process of plants, these are a
series of biological changes, the degree of which
reflects the growth speed. Sound has an important
* Corresponding author. Tel./fax: /86-23-6533-7319.
E-mail addresses: wangbc2000@yahoo.com.cn,
wangbc@cqu.edu.cn (W. Bochu).
0927-7765/03/$ - see front matter # 2003 Elsevier B.V. All rights reserved.
doi:10.1016/S0927-7765(03)00128-0
Whether the effect promotes or limits the growth
depends on the frequency, intensity and acting
time. At the same time, to advance the growth of
different plants there are different ‘best frequencies
and intensities’, which needs experiments to prove.
D.E. Ingber [8] suggests that it is very important
that it is not enough to depend on growth factors
in the growth of cell, whose biological pattern is
always decided by mechanical environment,
though the cell’s form is controlled by chemical
factors. Furthermore, the pathbreaker in biome-
chanics field, Yuanzhen Feng pointed out that the
relation between mechanics and growth is the soul
of biomechanics. So is in plant biomechanics.
It has been demonstrated that stimulation such
as sound field, supersonic, electromagnetic field,
30 W. Bochu et al. / Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34
micro gravity and mechanic vibration have effect
on plants [9
/11].
The use of supersonic in bioengineering and
pointed out that supersonic with low strength
could improve activity two times [12].
It is reported in Yiyao Liu’s article that sound
field stimulation has double effect including limit
and promotion on chrysanthemum’s injured tis-
sue. He has found that the best intensity was 100
dB and the best frequency was 800 Hz. Given, the
condition, sound field stimulation can promote the
injured tissue’s growth conspicuously.
In the research on the effect of sound field
stimulation on Chinese goosebeery’s injured tissue,
Xiaocheng Yang reported that the best intensity
was 100 dB and the best frequency was 1000 Hz,
which could promote the plant’s growth notably.
Researches on gene level have been done, which
have proved that the biological function of plants
is related with gene expression. Until now, five
kinds of interrelated genes have been found:
TCH1, TCH2, TCH3, TCH4, TCH5 [13
/15].
In this article we designed an apparatus to
stimulate the plant and investigated. Then we
fresh weight of other group  fresh weight of control group
fresh weight of control group
observe the biological effects of sound field
stimulation on paddy rice seeds, and explored the
mechanism of sound action initiatively.
2. Material and method
In this experiment in which single factor is
changed, paddy rice seeds from the same series
are cultivated after 12 h immersion. They are
divided into two groups of five teams, which
contain 50 drops of the seed each team. There is
one team without stimulation as control team in
both groups. In the group 1, seeds are stimulated
under 400 Hz with different intensities (96, 101,
106, and 111 dB). In the group 2, seeds are
stimulated under 106 dB with different frequencies
(200, 400 Hz, 1 and 4 kHz). Each group was
stimulated twice a day (30 min one time) for 2 days
and then put into steady-temperature and lighting
cultivation-tank under 25 8C for 5 days.
2.1. Determination of the germination index, height
of stem, relative increase rate of fresh weight,
number of roots and total length of root system
According to the number of seeds which has
germinated every day; the germination was calcu-
lated [16]. After 7 days cultivation, 15 drops of
seeds from each team were selected to measure the
height of stem, relative increase rate of fresh
weight, number of roots and total length of root
system directly.
relative increase rate of fresh weight%
Mi  Mck
 100%
Mck
100%
2.2. Determination of the activity of root system
[17]
TTC is a H-receptor, which is functioned by
dehydrogenase whose activity is due mutual to the
seed’s activity. The activity of TTC is evaluated
from the mass of TTC in per gram root [18].
2.3. Determination of penetrability of membrane
Cell membrane is a kind of selective penetrating
membrane, which can be injured in terrible envir-
onments, e.g. utmost temperature, salting and air
pollution. The injury could promote its penetr-
ability and let out electrolyte, and the conductivity
 W. Bochu et al. / Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34 31

Table 1
Biological effects of paddy rice under given sound field frequency

Intensity of sound field

CK 96 dB 101 dB 106 dB 111 dB

Germination index (drops/day) 48.49/0.04 48.29/0.03 48.59/0.04* 48.69/0.05* 48.39/0.03

Increase rate of fresh weight (%) 0.859/0.03 1.289/0.04* 1.519/0.05* 1.349/0.03
Plant height (cm) 27.59/1.4 29.89/1.3 30.39/1.5 32.29/1.5** 31.59/1.4
Number of root 4.69/0.7 4.49/0.6 4.69/0.9* 5.19/0.7** 4.89/0.7
Total length of roots (cm) 5.979/0.87 6.139/0.85 6.759/0.92** 7.819/1.29** 7.069/0.95*
Activity of roots (TTC mg/gFW) 1.0259/0.08 1.1259/0.13* 1.1839/0.11 1.2649/0.05** 1.2189/0.11*
Penetrability of cell membrane (%) 70.469/2.13 71.329/1.98 65.239/2.84* 61.269/2.33** 63.189/1.63

*P B/0.05, **P B/0.01. *The sound field frequency was 400 Hz.

can be measured by DDS-II */an apparatus for
conductance mensurating [19].
3. Results
3.1. The biological effects of different sound field
intensities on paddy rice
Table 1.
3.2. The biological effect of different sound field
frequencies
Table 2.
The results also showed that the number of
leaves, plant height, relative increase rate of fresh
weight and rooting ability are markedly enhanced
under the sound of 106, and 400 Hz, suggesting
that suitable sound stimulation is beneficial for the
absorption of nutrition and the transformation of
energy, so as to accelerate the growth of paddy
rice. That possibly causes the improvement of
protein content. At the same time, it is good for
the root system and efficiency of breeding.
But excessive sound stimulation limits the ger-
mination and growth of seed. When the frequency
reaches 4 KHz and intensity surpasses 111 dB, it
becomes a kind of mechanical injury (Figs. 1
/6).
Proper sound stimulation (about 400 Hz, 106
dB) is advantageous to reduce the penetrability of
cell membrane(Fig. 7). It can improve the protec-
tive ability of paddy rice in unfavorable environ-
ments. When the frequency and intensity exceed
the given range, the penetrability of membrane will
extend. It is proved that the sound of 400 Hz can
improve the float of cell membrane, strengthen the
mutual function between membrane lipo and
membrane protein. As a result, the growth of
plant is improved.

Table 2
Biological effects of different sound field frequencies on paddy rice

Frequency of sound field

CK 200 Hz 400 Hz 1 kHz 4 KHz

Germination index (drops/day) 48.49/0.03 48.69/0.05 48.59/0.04* 48.39/0.05* 48.69/0.04

Increase rate of fresh weight (%) 1.139/0.04 1.529/0.05* 1.079/0.03* /0.239/0.02
Plant height(cm) 33.79/1.3 35.29/1.8 38.49/2.3** 36.59/1.8* 32.19/1.2
Number of root 4.89/0.8 5.39/0.9* 5.89/1.0** 5.49/0.9* 4.39/0.6
Total length of roots (cm) 6.459/1.12 7.329/0.92 8.049/1.15** 7.559/0.93** 6.229/1.02*
Activity of roots (TTC mg/gFW) 1.0779/0.068 1.1939/0.105* 1.2529/0.032** 1.2019/0.078 1.0949/0.097
Penetrability of cell membrane (%) 69.219/2.42 64.059/2.12** 62.599/1.83** 67.289/2.35* 70.579/2.25

*P B/0.05, **P B/0.01. *The sound field intensity was 106 dB.
32 W. Bochu et al. / Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34
Fig. 1. The Effect of intensity and frequency on the germina-
tion index.
4. Discussion
Under the sound wave of 200 and 400 Hz, the
height of stem, fresh weight, number of root, total
length of root all rise evidently while the penetr-
ability of cell membrane drops. When the fre-
quency achieves 1 KHz, the data above begin to
change to the opposite direction. With the stimu-
lation frequency of 4 KHz, it shows obvious
injury. Similarly, when we change the sound
intensity in certain range under 400 Hz, it ex-
presses beneficial effect. If the intensity goes
beyond normal limit, it causes reverse effect.
It is thought that 400 Hz is outstandingly
meaningful in the stimulation of sound field.
Because it has important effect on the action of
cell cliff, the secondary construction of membrane
protein and the floating of cell membrane [1
/3]. It
is inferred that only when the frequency of the
stress coincides with the biological frequency of
cell itself, does the sound stimulation have its
Fig. 2. Effect of intensity and frequency on the increase rate of
fresh weight.
Fig. 3. Effect of intensity and frequency on the plant height/
stem.
function. In this experiment we also find that 400
Hz and 106 dB are the ‘best frequency and
intensity’ to paddy rice seed.
All the materials chosen in this experiment are
the same plump paddy rice seed. If we used old
seeds, we think the sound stimulation may have
effect on two quotas. Because many researchers
[4,20] believed that certain sound stimulation can
accelerate the activation of SOD and POD and cut
down the IAA activation and the content of
MDA. SOD and POD are the oxidizing system.
It can clear the free base produced by ultra
oxidization and repair the injured cell membrane
at some increase of IAA level, which could speed
up the plants growth, MDA is produced by the
membrane lipo ultra oxidization. Its content
expresses the degree of ultraoxidization. So it is
reasonable that if old seed is processed by certain
sound stimulation, the germination index will be
developed. Moreover, we find that sound stimula-
tion might enhance the germination speed.
Why sound stimulation improves the fresh
weight and height of plant? We believe that there
are three causes. First, the sound field influences
the cell cycle of paddy rice cell and speeds up its
reproduction rate. Second, the sound field trans-
fers energy into the cell in the form of field and
drives the floating of nutrition. Third, it also
affects the membrane materials to cause changes
of biological function of membrane and enhance
the metabolism of cells.
There are many important indexes to judge the
growth state of plant, such as rooting ability and
activity of root. They affect directly the root’s
 W. Bochu et al. / Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34
Fig. 4. Effect of intensity and frequency on the number of root.
function, which include the composition of some
biological active material and abruption of chemi-
cals and water. So the study on root has much
significance in raising seedlings in factory. Here is
a mechanism that sound field has connection with
the density of Ca2 which is the second messenger
in the message delivery of cell. It has been proved
that Ca2 participates in the process of root
growth. Sun Day [14,21] has ever reported that
the lack of Ca2 in the root of wheat seedling
wound cause consist ant fall of the root growth.
Evidently, it amply indicates that Ca2 has an
effect on the activity of root. The experiment also
demonstrates the importance of Ca2 in the
growth of root [5,14].
Cell membrane is the structure that is most
sensitive to the environmental stimulations. The
structure is made up of protein, lipo and chemical
conveyance, and it contains message delivery and
Fig. 5. Effect of intensity and frequency on the total length of
roots.
33
Fig. 6. Effect of intensity and frequency on the activity of
roots.
the metabolism process of protein and enzyme.
Cell membrane is a type of selective membrane. Its
penetrability influences the plant resistance against
harmful matters and endurance in terrible envir-
onment.
Sound field stimulation with certain frequency
and intensity could repair the injury made in
adverse circumstance to some extent. It can also
promote the activity of the enzyme system of
metabolism, keep the metabolism and function of
protoplast steady [4,6,7], reduce the penetration
and then strengthen its resistance. We believe that
promoting the membrane’s selective penetration
can suitably enhance the conveyance of materials
and improve the cell’s growth. It is perceived that
sound field stimulation with certain frequency and
intensity can indeed strengthen some biological
indexes of plants and accelerate the growth of
them.
Fig. 7. Effect of intensity and frequency on the penetrability of
cell membrane.
34 W. Bochu et al. / Colloids and Surfaces B: Biointerfaces 32 (2003) 29
/34
Acknowledgements
This paper was supported by Chongqing Science
and Technology Committee.
References
[1] B.C. Wang, A. Yoshikoshi, A. Sskanishi, Carrot cell
growth response in a stimulated ultrasonic environment,
Colloids Surf. 12 (1998) 89
/95.
[2] B.C. Wang, H.C. Zhao, Y.Y. Liu, The effects of alter-
native stress on the cell membrane deformability of
Chrysanthemum cells, Colloids Surf. B: Biointerf. 20
(2001) 321
/325.
[3] X.C. Yang, B.C. Wang, C.R. Duan, C.Y. Dai, Biological
effects of actidiniachinensis callus on mechanical vibration,
Colloids Surf. B: Biointerf. 25 (2002) 197
/203.
[4] Y.Y. Liu, B.C. Wang, H.C. Zhao, Alternative stress effects
on the Ca2 distribution in the chrysanthemum callus
cells, Colloids Surf. B: Biointerf. 22 (2001) 245
/249.
[5] Y.Y. Liu, B.C. Wang, X.F. Long, C.R. Duan, A.
Sakanishi, Effects of sound field on the growth of
Chrysanthemum callus, Colloids Surf. B: Biointerf. 24
(2002) 321
/326.
[6] Y. Jia, B.C. Wang, X.J. Wang, C.R. Duan, X.C. Yang,
Effect of sound stimulation on roots growth and plasma-
lemma H  -ATPase activity of chrysanthemum (Gerbera
jamesonii ), Colloids Surf. B: Biointerf. 27 (2003) 65
/69.
[7] H.C. Zhao, B.C. Wang, Y.Y. Liu, Influence of water stress
on the lipid physical state of plasma membranes from P.
Betuloefocia. Bqe leaves, Colloids Surf. B: Biointerf. 19
(2000) 181
/185.
[8] D. Ingber, Integrins as mechanochemical transducers,
Curr. Opin. Cell Biol. 3 (1991) 841
/848.
[9] D.S. Bush, Calcium regulation in plant cells and its role in
signaling, Annu. Rev. Plant Physiol., Plant Mol. Biol. 46
(1995) 95
/122.
[10] L.N. Fraze, D. Moor, The role of calcium accumulation
and cytosketetal elements in perception and response of
coprinus cinerous to gravity, CQSPAR’94, Abstracts,
1994, 280.
[11] S.T. Roux, Calcium as a mediation of plants directional
grown response to gravity, in: Fundamentals of Space
Biology, Japan Science Society Press, 1990, pp. 57
/67.
[12] S.Y. Qiu, R.H. Yao, M.H. Zong, The appliance of
supersonic in bioengineering, Dev. Bioeng. 3 (1999) 45
/48.
[13] M.J. Jaffe, Thigmomorphogenesis: the effect of mechanical
perturbation on plant, Plant Growth Regul. 3 (1993) 313
/
324.
[14] J. Braam, W. Davis, Rain, wind and touch induced
expression of calmodulin and related gene in Arabidopsis,
Cell 60 (1990) 357
/364.
[15] J. Braam, Arabidopsis TCH4 regulated by hormones and
the environment encodes axyloglancan endotransglycosy-
lase, Plant Cell 7 (1995) 1555
/1556.
[16] Z.H. Gu, Inquiry on method of seed vigour test: physio-
logic measure of seed germination, Seed 3 (1982) 11
/16.
[17] J. Bramm, Regulated expression if the calmodulin-related
TCHs in cultured Arabidopsis cells induction by calcium
an heat shock, Proc. Natl. Acad. Sci. USA 98 (1992) 3213
/
3216.
[18] B. Bai, J. Jin, Improvement of TTC methods for maize
root, Maize Sci. 2 (1994) 44
/47.
[19] H.K. Lichtenthaler, Annu. New York Acad. Sci. 851
(1998) 187
/198.
[20] S. Akers, C. Mitchell, Seismic stress effect on vegetative
and reproductive development of Alaska pea cell, J. Bot.
62 (1984) 2011
/2015.
[21] D.S. Bush, Calcium regulation in plant cell and its role in
signaling, Annu. Rev. Plant Physiol. Plant Mol. Biol. 46
(1995) 95
/122.