The Effect of Audible Sound on the Germination and Root Elongation of Selected Seedlings

Western Michigan University Western Michigan University

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Masters Theses Graduate College

4-1975

The Effect of Audible Sound on the Germination and Root The Effect of Audible Sound on the Germination and Root

Elongation of Selected Seedlings Elongation of Selected Seedlings

Micheal Dennis Walton

Western Michigan University

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Seedlings" (1975).

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THE EFrriCT OF AUDIBLE SOUND ON THE GERMINATION AND

ROOT ELONGATION OF SELECTED SEEDLINGS

Micheal Dermis Walton

A Thesis

Submitted to the

Faculty of The Graduate College

in partial fulfillment

of the

Degree of Master of Arts

Western Michigan University

Kalamazoo, Michigan

April 1975

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ACKNOWLEDGEMENTS

I' am very grateful for the advice and assistance I received

in the undertaking of this project to Dr. Leo C. Vander 3eek and

the other members of my thesis committee, Dr. Richard Y,\ Pippen

and Dr. Eugene 3emstein.

A- special thanks to a good friend iir. F e m e (3ud) Ellis

without whose help this task would have been much more difficult.

His assistance in the setting up of the experiment and in running

the data through the computer was of immeasurable value.

Finally, a word of thanks to Mr. Parrish, who works in the

electronics laboratory, for his help in repairing and replacing

some of the sound equipment which was necessary before the experiment

could be started.

Micheal Dennis Walton

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MASTERS THESIS

M-7033

WALTON, Micheal Dennis

THE EFFECT OF AUDIBLE SOUND ON THE GERMINATION

AND ROOT ELONGATION OF SELECTED SEEDLINGS.

Western Michigan University, M.A., 1975

Biology

Xerox University Microfilms , Ann Arbor, Michigan 48106

T H IS D IS S E R T A T IO N H A S R EFN M IC R O F IL M E D F X A C T I Y AS R F O F IY /F D

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TABLE OF CONTENTS

CHAPTER

i n

P’ACki

INTRODUCTION

...............................

LITERATURE RSVISV;

...........................

LETHODS AND MATERIALS

........................

Plant Material

............................

Sound Source

..............................

Growth Conditions

.........................

Determination of Germination and Growth

......

Statistical Analysis

.......................

RES U LT S

....................................

Root Elongation

...........................

Germination

...............................

DISCUSSION

.................

...............

iii

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LIST C? TABLr.3

TA3LB

1 lie an length in as of longest root of

germinated cucumbers for control and

sound treated seeds

...........................

2 iiiean length in as of longest root of

germinated oats for control and sound

treated seeds

..........................

3 mean length in ms of longest root of

germinated wheat for control and sound

treated seeds

................................

1 Per cent germination of cucumber for control

and sound treated seeds

.......................

5 Per cent germination of oats for control

and sound treated seeds

.......................

6 Per cent germination of wheat for control

and sound treated seeds

........................

7 Two-way analysis of variance (unbalanced

case) for root elongation data of

cucumbers for runs 1 and 2

.....................

8 Two-way analysis of variance (unbalanced

case) for root elongation data of

cucumbers runs 3 and 1

............

9 Two-way analysis of variance (unbalanced

case) for root elongation data of

cucumbers runs 5 and 6

........................

1C Two-way analysis of variance (unbalanced

case) for root elongation data of

cucumbers runs 7 and 8

........................

11 Two-way analysis of variance (unbalanced

case) for root elongation data of

cucumbers all runs

............................

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TABLE PAGE

12 Two-way analysis of variance (unbalanced

casej for root elongation data of

oats runs 1 and 2

............................

13 Two-way analysis of variance (unbalanced

case) for root elongation data of

oats runs 3 and 4

............................

14 Two-way analysis of variance (unbalanced

case) for root elongation data of

oats runs 5 and 6

............................

15 Two-way analysis of variance (unbalanced

case) for root elongation data of

oats runs 7 and S

............................

16 Two-way analysis of variance (unbalanced

case) for root elongation data of

oats all runs

................................

17 Two-way analysis of variance (unbalanced

case) for root elongation data of

wheat runs 1 and 2

...........................

18 Two-way analysis of variance (unbalanced

case) for root elongation data of

wheat runs 3 ana 4

...........................

19 Two-way analysis of variance (unbalanced

case) for root elongation data of

wheat runs 5 and 6

...........................

20 Two-way analysis of variance (unbalanced

case) for root elongation data of

wheat runs 7 and 8

.............

21 Two-way analysis of variance (unbalanced

case) for root elongation data of

wheat all runs

...............................

22 Two-way analysis of variance of cucumber

germination data

.............................

23 Two-way analysis of variance of oat

germination data

..........

24 Two-way analysis of variance of wheat

germination data

.............................

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THE EFFECT OF AUDIBLE SOUND ON THE GERMINATION AND

•ROOT ELONGATION OF SELECTED SEEDLINGS

Introduction

Sound is energy which travels in the fora of sound waves. It is

present throughout the environment so that all living organisms are

exposed to it. lian can detect sounds ranging in intensity from 10“^

watts/m^ to 1 watt/m^. The lower intensity is barely audible while

the higher intensity is pain producing. The range of frequencies

which man can detect is referred to as the audible sound range. Those

sounds of frequencies above the audible range are called ultrasounds.

The frequency of sound is measured in Hertz or cycles per second

and the intensity of sound is expressed in decibels (db). The decibel

is defined according to a logarthmic scale. The intensity level in

decibels r 10 log l/l0 where ID is the intensity of the faintest

audible sound which is 1 0 " ^ watts/m^. This being the case, the

intensity level of the least audible sound is 10 log (10_*^/lC~-^)

which is zero.

kost experiments reported in the literature involve the use of

ultrasound (sound of higher frequency then audible sound). Ultra

sound has very destructive effects on living cells. Yeast and bacterial

cells present in milk have been destroyed upon exposure to ultrasonic

waves (Beckwith and 7/eaver, 1936). The destructive effects of ultra

sound are largely due to cavitation. Cavitation involves the formation

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of partial vacuums -within the cell due to the separation of intra

cellular structures (Clark and Hill, 1969). Ultrasonic waves are

frequently used in the destruction of microbial cells and the

separation of intra-cellular particles (El'Piner, 1964).

Experiments with audible sound reported in the literature

generally involved animals. Disruption of animal cells by audible

sound may be produced by the explosion of the organism as a result

of the internal release of dissolved gases, or by actual tearing

apart of tissues as a result of rapid alterations of tension and

compression produced in the surrounding medium by the vibrations

(Chambers and Gaines, 1932). Plant experimentation with audible

sound generally concerns effects on germination and inhibition

or stimulation of plant growth.

This work was initiated to examine the effects of audible

sound on the germination and growth of cucumbers, oats, and wheat.

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LITERATURE REVIEW

The effects of audible sound on animals and animal cells has

been investigated in several ways. Chambers and Gaines (1932) found

that the water flea, Daphnia pulex, when exposed to sonic irradiation

in a shell vial filled with water, was killed within one second. The

exoskeleton was shattered, the soft body parts were churned into an

amorphous pulp, and clouds of colloidal material were released into

the surrounding water. 3rys’nenskii and Meskhaeleva (1969) found that

animals subjected to the prolonged action of sound exhibited a lower

coagulation activity of the blood. The sound weakens the defense

reactions which develope after the injection of thrombin. The death

rate amoung the experimental animals was 72.5% while the death rate

for the control animals was 40.9*. Aleksandrovskaya and Chezhenkova

(1970) found that a weak sound (200 c.p.s.) in the rabbit cerebral

cortex enhanced the slow waves and spindles, particularly in the motor

area. In the deep layers of the motor area, the weak sound brought

about an increase in astrocytes. In the deep layers of the projection

area (auditory cortex) the astrocytes decreased in number.

Xoitchev (1969) found that there was an increase of sodium in

the organ of Corti after exposure to sound. This was attributed to

changes in the permeability of excited membranes. Chambers and Harvey

(1931) found that tadpoles of 3ufo punctata when held in shell vials

filled with water, above the vibrator, were killed within one minute.

After one minute of sound treatment, bubbles of gas could be seen in

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the abdominal cavity, and after two minutes the tadpoles actually

exploded. Small fish were killed in three to four minutes and

immature frogs were killed in about ten minutes.

Most of the work on the effects of audible sound on plants

and plant cells concerns growth and germination. Northen and

hacVicar (1939) showed that sound lowered the elasticity of the

cytoplasm of Spirogyra by 20 per cent to 88 per cent. This could

be seen by the displacement of chloroplasts in sound treated

Spirogyra filaments. Singh (1959) reported that the water plant

Hydrilla verticillata had an increased rate of photosynthesis

after exposure to sound waves from an electric bell. 3oth the rate

and the total volume of oxygen evolved from sound-excited Hydrilla

plants were 60 per cent to 100 per cent higher then the control

plants. Gnanam (1959) observed an increase in the photosynthetic

rate of Spirogyra when exposed to sound. The volume of oxygen

evolved in excited plants vras much higher then the control,

showing thereby, that in sound excited plants synthesis and

production of food was higher.

Some rather non-scientific literature has been concerned

with the effect of music on plants. RetaHack (1973) found that

acid rock, after being played to plants for five days, had very

devastating effects on plant growth. She observed a bending of the

plants away from the speaker and a decrease in root developement.

liusic played by Ravi Shankar had no effect on root developement

and the plants were found to be bending toward the speaker. Some

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reference to the effects of music on plants has been recorded in

scientific literature. Ponniah (1958) played a single note to

Mimosa pudica, Impatiens balsamina, Tagetes erecta, and Hydrilla

verticillataa She found an increase in root developement, total

number of leaves, and length of branches. Singh and Ponniah (1955)

found that the musical sound of the Veena stimulated the growth

of balsam plants in all parameters measured.

Lisenkov (1966) found that seeds of Siberian larch exposed

to sound treated water had an increase in ground germination,

increased frost resistance, and better growth. The enzyme activity

of the germinated seeds was also increased under certain conditions.

Amylase activity increased when the seeds were soaked in sound

treated water for periods between 0.5 and 1.0 hours.

Weinberger and Measures (1968) exposed spring and winter

wheats (Triticum aestivum) to a single audible frequency of

sound at a given time. They found better germination and an

increase in growth in winter wheat (var. Rideau) and in some

cases this was temperature or frequency dependent. Spring wheat

(var. Marquis) showed an increase in germination at 2 degrees C

and 10 degrees C, but not at 25 degrees C. The growth of spring

wheat was found to vary with different treatments. It was suggested

that the increase in germination at 2 degrees C and 10 degrees C

may involve the production of ethylene.

The same workers in another experiment using spring wheat

found a significant increase in some of the parameters measured

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when 3^0 Hz and 5,000 Hz were used. However, plant growth was not

significantly stimulated by treatment with either 1,250 Hz or

12.000 Hz sound frequencies (keasures and Weinberger, 1970)•

Some negative effects of sound on growth have also been

reported in scientific literature. Weinberger and Das (1972)

found that continuous exposure of Scenedesmus obtusiuscuius to

1.000 Hz resulted in a decrease in the rate of cell division. A

decline of 15 per cent was obtained in 18 hours. The normal rate

of cell division did not return until two life cycles had elapsed.

The Scenedesmus cells were most sensitive to sonic shock in the

early part of their life cycle. Yfoodlief et al., (1969) also found

some negative effects of sound on plants. Tobacco plants subjected

to random noise showed a decline in growth rate of over 10 per cent.

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METHODS AND MATERIALS

Plant Material

The three types of certified seeds used in these experiments,

wheat (Triticua aestivum), oats (Avena sativa var. AuSable), and

cucumber (Cucumis sativis var. marketer) were obtained from Farm

Bureau Services Inc., Kalamazoo, Michigan. VJhen not being used for

experimentation all seeds were stored in a refrigerator at approx

imately 8 degrees C.

Sound Source

Sound at a frequency of 600 Hz was generated by an audio-

oscillator^- connected to an amplifier^. Sound intensity was

measured by using a sound level meter3 using the 'G' weight. The

audio-oscillator was adjusted to give sound of the intensity of

102 plus or minus 3 decibels in each of the two growth chambers.

In all experiments a nine inch speaker was used. The speaker was

suspended by heavy strings near the center of the growth chamber.

Growth Conditions

Both control series and experimental series consisted of

^-Audio-oscillator made by Hewlett Packard Co., Pala Alto,

California (Model 200 CD).

^Amplifier made by the David Bogen Co., New York, New York

(jiodex m l j.

3Sound level meter made by the General Radio Co., Nest

Concord, Massachusetts.

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5 plastic dishes-*- (155 on by 63 mm). Each dish was lined with Armstrong

ft6 filter paper^ and was moistened with 15 ml of distilled water. A

total of 20 seeds were randomly scattered in each dish giving 100

seeds for each run. The plastic containers were then placed in clear

polyethylene bags3 which were closed by using wire twists. After ail

dishes were moistened and planted, they were placed in Sherer Controlled

Environment Chambers^-. Experiments in which cucumbers were tested and

those in which oats were tested were run simultaneously. The experiments

with wheat were run separately since there was not enough room in

the growth chamber to run 3 sets of dishes at the same time.

The dishes were placed near the center of the growth chamber

in a circular pattern. The growth chambers (experimental and control)

were kept in constant darkness at a temperature of 20-21 degrees C.

The experimental series was continuously exposed to sound. The control

series was ran in a second growth chamber under exactly the same

conditions and at the same time. The only sound in the control

chamber was due to normal background noise. After 5 days all seeds

■were removed and measured. A total of 8 separate runs, 4 replications

^Plastic containers were obtained from 3radley Industries,

Inc., Franklin Park, Illinois.

2?aper manufactured by the Armstrong Cork Co., Lancaster,

Pennsylvania.

^3ags made by Union Carbide Corp., Consumer Products Division,

270 Park Avenue, New York, New York, and sold under the trade name

of 'Glad*.

^Sherer-Gillett Co., Marshall, Michigan.

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of the experiment, were made for each of the 3 types of seeds.

The growth chambers were interchanged for each succeeding run

sc that the 1 experimental' chamber became the 'control1 chamber

on the next run.

Determination of Germination and Growth

Any seed was considered to have germinated if its root reached

a length of 1 mm or longer. Growth was determined by using the

method of Thompson et al., (1915) where the longest root of each

germinated seed was measured in mm. A mean root length was determined

for both control and experimental groups by using only root lengths

of seeds that did germinate.

Statistical Analysis

Data on root elongation were analyzed by means of library,

program # 1.9.2. using Western Michigan Universities PDP-10

computer. This program is a two-way analysis of variance (unbalanced

case) where factor 1 is the difference between control and experimental

treatments and factor 2 is the .difference between rims. Seeds that

did not germinate were not included in the analysis. Data on

germination were analyzed using library program# 1.9.1. (version 2).

This program is also a two-way analysis of variance. It shows the

significance of control/experimental, replication, and growth chamber

differences. It does not consider within cell variability. In both

programs used a probability level was calculated from the 'F' value.

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RE SU LTS

Root Elongation

Tables 1, 2, and pages 13, 14, and 15 show the average length

of the longest root of germinated cucumber, oat, and wheat seeds under

both control and experimental conditions. In the case of cucumbers and

oats the grand mean (GM) of the control group is significantly greater

then the grand mean of the experimental group. In the case of wheat the

grand mean of the control group is not significantly greater then the

grand mean of the experimental group. Tables 7-21 on pages 19-33 show

the results of statistical analysis.

Cucumber seeds

YVith cucumbers (see Table 1, page 13), the grand mean of the

control group is 56.61 mm while the grand mean of the experimental

group is 47.09 The difference observed is significant at the 0.1

per cent level (see Table 11, page 23). A significant difference (at

the 3.5 per cent level) is found between the 8 runs of the experiment.

Interaction (which determines if differences are due to experimental

conditions or some other factors) is significant at above the 23 per

cent level. Since interaction is not significant at below the 5 per

cent level, no further analysis of root elongation data for cucumbers

is needed.

Oat seeds

The data for oats (see Table 2, page 14) is similar to that

for cucumber showing an inhibition of growth in the sound treated

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•group. The grand mean of the control group is 112.17 mm and the grand

mean of the experimental- group is 99.23 am. The difference between

the two is a significant difference at below the 0.1 per cent level

(Table 16, page 28). There is a significant difference between the

8 runs (below the 0.1 per cent level). The interaction is not

significant being above the 70 per cent level. Once again, since

interaction is not significant no further analysis of root elongation

data for oats is needed.

Wheat seeds

The data for wheat (see Table 3, page 15) are different than

the data for cucumbers and oats showing no real inhibition of growth

in the sound treated group. The grand mean of the control group is

105.03 mm and the grand mean of the experimental group is 101.61 mm.

The difference between the two is not significant (Table 19, page 31).

There is a significant difference between the 8 runs (below the 1.1

per cent level). The interaction is significant only above the 8.5

per cent level. Since the difference between the control and sound

treated -wheat seeds is not significant no further analysis of the

data is required.

Germination

Tables 1, 5, and 6; pages 16, 17, and 18 show the per cent

germination for cucumbers, oats, and wheat. Tables 22, 23, and 21;

pages 31, 35, and 36 show the level of significance for-the differences.

In cucumbers, oats, and wheat there is no significant difference

in per cent germination for either growth chambers, replications,

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or control/experimental differences.

Cucumber seeds

The data indicate that there is no significant difference in

per cent germination between control and experimental plants (22.1

per cent level), between replications (36.5 per cent level), or

between growth chambers A and B (22.1 per cent level).

Oat seeds

The data indicate that there is no significant difference in

per cent germination between control and experimental plants (84.4

per cent level), between replications (8.0 per cent level), or

between growth chambers A and B (55.5 per cent level).

VTheat seeds

As in cucumbers and oats, the data indicate that there is no

significant difference in per cent germination between control and

experimental plants (18.5 per cent level), between replications

(18.5 per cent level), or between growth chambers A and 3 (42.5

per cent level).

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Replications

G il

56.24

56.61 62.96

59.08

47.95

Control

53.81

51.85

52.11

38.31

Sound

Runs

Table 1. iiean length in rnrn of longest root of germinated cucumbers for control

and sound treated seeds. GM = Grand Mean. (Significant to 0.1 mm).

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Control

Sound

Table 2.

Replications

GM 1

112.17 95.47

119.94

109.51

104.14

107.78

121.53

122.85

116.11

99.23

83.97 104-93

98.56

86.78

93.74 108.97

108.95

107.93

1 2

Runs

Mean length in mm of longest root of germinated oats for control and

sound treated seeds. GM = Grand Mean. (Significant to 0.1 mm).

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Replications

Control

105.03

107.04 107.76

106.15

100.62

100.85

102.79

108.78

106.26

Sound

104.61

105.53 109.33

97.35

107.90

96.88

107.86

102.52 109.52

.......

Runs

Table 3« Mean length in ram of longest root of germinated wheat for control and

sound treated seeds. GM = Grand Mean. (Significant to 0.1 nun)

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Table 5. Per cent gemination of oats for control and sound treated seeds.

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Grand mean for control

Grand mean for experimental

56.61 mm

47*09 nun

FACTOR

F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

20.74

0.000

Difference between runs

44.28

0.000

Interaction

1.73

0.190

Table 7. Two-way analysis of variance (unbalanced case)

for root elongation data of cucumbers

(library program jj 1.9.2) for runs 1 and 2.

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Grand mean for control

Grand mean for experimental

56.61 mm

47.09 mm

FACTOR

F VALUE

PROBABILITY

Weighted means analysis of variance

Pifference between control/experimental

48.52

0.000

Difference between runs

7,88

0.005

Interaction

1.20

0.275

Table 8 . Two-way analysis of variance (unbalanced case) for root elongation data of cucumbers

(library program // 1 .9 *2) for runs 3 and 4 .

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Grand mean for control

Grand mean for experimental

56.61 nan

47.09 mm

FACTOR

F VALUE P R O B A B IL IT Y

Weighted means analysis of variance

Difference between control/experimental

11.92

0.001

Difference between runs

16.18

0.000

Interaction

2.70

0.101

Tabl e 9. T w o - w a y a nalysis of varian ce (u n b a l a n c e d case) fo r root e l o n g a t i o n data of c ucum be rs

(libr ary p r o g r a m // 1.9.2) f o r runs 5 and 6.

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Grand mean for control

Grand wean for experimental

5o.6i, mm

47.09 hum

FACTOR F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

13.51

0.000

Difference between rims

1.72

0.191

interaction

0.18

0.668

Table 10. Two-.vay analysis of variance (unbalanced case) for root elongation data of cucumbers

(library program y 1 ,9.2 ) for runs 7 and 8 .

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Grand mean for control.

Grand mean for experimental

56.61 mm

47.09 mm

FACTOR F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

Difference between runs

Interaction

36.66

0.000

A.75

0.035

1.47

0.232

Table 11. Two-way analysis of variance (unbalanced case) for root elongation data of cucumbers

(library program ft 1 .9 .2 ) all runs.

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Grand mean for control 112.17 jnm

Grand mean for experimental 99*23 inm

FACTOR F VALUE PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

•15.41

0.000

Difference between runs

45*27

0.000

Interaction

0.27

0.604

Table 12. Two-way analysis of variance (unbalanced case) for root elongation data of oats

(library program if 1 .9*2 ) for runs 1 and 2 .

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Grand mean for control

Grand mean for experimental

112.17 mm

99.23 mm

FACTOR

F VALUE PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

13.19

0.000

Difference between runs

4.83

0.029

Interaction

0.68

0 . 4 H

Table 13. Two-way analysis of variance (unbalanced case) for root elongation data of oats

(library program # 1 .9 .2 ) for runs 3 and 4 .

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Grand mean for control

112,17 mm

Grand mean for experimental

99*23 mm

FACTOR F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

17.91

0.000

Difference between runs

21.25

0.000

Interaction

0.05 0.815

Table 1^. Two-way analysis of variance (unbalanced case)

for root elongation data

of oats

(library program ft 1 .9 .2 ) for runs 5 and 6 .

O '

Grand mean for control

Grand mean for experimental

112.17 mm

99.23 mm

FACTOR F VALUE P R O B A B IL IT Y

Weighted means analysis of variance

Difference between control/experimental

9.74

0.002

Difference between runs

1.20

0.274

Interaction 0.66

0.419

Ta b l e 15. T w o - w a y a nalys i s of v ari a n c e ( u nb al a n c e d case) f o r r o ot e l o n g a t i o n d a ta of oats

(l i brar y p r o g r a m

1. 9. 2) f o r runs 7 a n d B.

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Grand mean for control 112.17 mm

Grand mean for experimental 99*23 mm

FACTOR F VALUE PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

32.05

0.000

Difference between runs

21.82

0.000

Interaction 0.15

0.702

T a b l e 16. Tw o - w a y an a l y s i s of v a r i a n c e ( u nb a l a nce d case) f or root e l on g a t i on d a ta of oats

(library p r o g r a m

1.9.2) all runs.

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Grand mean for control

Grand mean for experimental

105o03 mm

104.61 mm

FACTOR

F VALUE

P R O B A B IL IT Y

Weighted means analysis of variance

Difference between control/experimental

Difference between runs

Interaction

0.00

0.60

0.28

0 . 9 9 3

0.440

0.598

Table 17. Two-way analysis of variance (unbalanced case) for root elongation data of wheat

(library program // 1 .9 .2) for runs 1 and 2 .

v O

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Grand mean for control

Grand mean for experimental

FACTOR

105.03 mm

104.61 mm

•F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

0.09

0.767

Difference between runs

0.97

0.326

Interaction

9.92

0.002

Table 18. Two-way analysis of variance (unbalanced case)

for root elongation data

of wheat

(library program rf 1.9.2) for run3 3 and 4.

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'Grand mean for control 105.03 mm

Grand mean for experimental 104.61 mm

FACTOR F VALUE PROBABILITY

Weighted mean3 analysis of variance

Difference between control/experimental

0.04

0.832

Difference between runs

6.07 0.014

Interaction

2.97

0.085

Table 19. Two-way analysis of variance (imbalanced case) for root elongation data of wheat

(library program // 1.9.2) for runs 5 and 6.

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Grand mean for control

Grand mean for experimental

10^.03 mm

lOA.bl mm

FACTOR

F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

0.31

0.579

Difference between runs

O .69

0.408

Interaction 3.09 0.079

T a b l e 20. T w o - w a y analys is of v a r i a n c e (unb a lan c ed case) for root e l o n g a t i o n d a t a of wheat

( l ib r a r y p r o g r a m # 1.9.2) fo r runs 7 a n d 8.

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Grand mean for control

105.03 mm

Grand mean for experimental

10A.61 mm

FACTOR

F VALUE

PROBABILITY

Weighted means analysis of variance

Difference between control/experimental

0.17 0.683

Difference between runs 1 0.38

0.541

Interaction

0.00

1.000

Table 21. Two-way analysis of variance (unbalanced case)

for root elongation data

of wheat

(library program # 1.9.2) all runs.

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f a c t o r f v a l u e p r o b a b i l i t y

Cucumber

Difference between control/experimental

1.53

0.221

Difference

between replications

1.18

0.365

Difference

between chambers A and B

1.53

0.221

Table 22. Two-way analysis of variance of cucumber germination data (library program # 1.9*1

(version 2) ).

•t--

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FACTOR F VALUE PROBABILITY

Oats

Difference

between control/experimental

0.0/+

0.844

Difference between replications

3.17

0.080

Difference between chambers A and B

0.35 0.555

Table 23. Two-way analysis of variance of oat germination data (library program # 1.9.1

(version 2) ).

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FACTOR F VALUE PROBABILITY

Wheat

Difference between control/experimental

1.79 0.185

Difference between replications

1.79

0.185

Differences between chambers A and B

0.65 O.A25

Table 2k* Two-way analysis of variance of wheat germination data (library program // 1.9.1 (version

2) ).

VjJ

O '

DISCUSSION

Cucumbers and oats treated with 600 Hz audible sound showed a

significant inhibition of root elongation. Wheat did not show any

significant inhibition of root elongation. No significant change in

per cent germination was observed for any of the experimental plants.

In all runs, considerable variation is observed in both control

and experimental groups. In cucumbers, the control plants show a low

mean of 47.95 mm and a high mean of 62.96 mm: experimental plants

show a low mean of 38.31 mm and a high mean of 53*81 mm. In oats,

the control plants show a low mean of 95.47 mm and a high mean of

122.85 mm. The experimental plants show a low mean of 83.9? mm and a

high mean of 108.97 mm. Wheat control plants show a low mean of 100.62

mm and a high mean of 108.78 mm. Experimental wheat plants show a

low mean of 96.88 mm and a high mean of 109.52 mm.

Since these variations cannot be attributed to growth chamber

differences (see Tables 22, 23, and 24; pages 34* 35* and 36), some

other factors must be involved. It seems logical that morphological

variation in seeds may be a factor. Some selection in seeds was done

during the course of the experiment. For example, all seeds that

were obviously much larger or smaller then the 'average1 seeds were

not used in the experiment. Any broken or rotten seeds were discarded.

However, considerable variation in runs still occurred. Perhaps the

answer to this problem would be some type of screening apparatus. A

series of two screens could be placed one below the other so that

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very large seeds would not pass through the top screen. The seeds

collected on the bottom screen (which would have smaller openings

then the top screen) would be about the sane size with the very

snail seeds falling through both the first and second screens. It

also seems possible that size variations for runs would be smaller

if the seeds were allowed to grow for a longer period of time. It

was observed in other experimentation with Cyprus esculentus

(nutsedge) that tubers which were heavier and larger in size were

considerably larger then the lighter smaller tubers a few days

after germination. However, after several weeks of growth it was

observed that the differences in size were considerably less noticeabl

At 600 Hz of sound, treated cucumber and oat seeds showed a

significant inhibition of root elongation 'while wheat seeds did not.

Work previously done in this lab (Ellis, 1973) at 300 Hz sound

showed an inhibition of root elongation in wheat. However, it could

not be stated conclusively that the inhibition was due to sound

since interaction was significant. Experiments now in progress

(Gyimah) indicate that there is no significant inhibition of root

elongation in wheat, oats, or cucumber at 900 Hz or at 1,200 Hz.

Weinberger and Measures (1970) found an increase in growth in Marquis

spring -wheat at 300 Hz of sound. They found no effect on growth at

1,250 Hz of sound treatment. The differences observed in the growth

responses of different species of plants at different frequencies

of sound treatment is not disturbing. There are several possible

explanations for the occurrence of these varied responses.

In order for sound to effect growth, it seems logical that the

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sound waves must penetrate the seeds. Possibly the nake up of the seed

coat varies with different species. A thick seed coat would make it sore

difficult for sound waves to enter the seed then a thin seed coat. It

would be of interest to pursue this in further experimentation. It is

also possible that there are different biochemical pathways in different

species of plants. Some of these pathways may be disrupted by sound

waves while others may be unaffected. It right be that an enzyme or

an intermediate metabolic product could be broken down which would

result in the destruction of a particular pathway necessary for normal

growth to occur. Within the same plant species, it is quite possible

that only a specific frequency of sound would have the ability to

disrupt a particular metabolic pathway. It ’would be useful to carry

on further experimentation in which the amino acid levels of sound

treated and control seeds could be measured to see if there are any

significant differences.

Another interesting question is how do sound waves inhibit growth

in plants when this is seen to occur. Many investigators have attributed

the destructive effects of sound waves on cells to cavitation (Chambers

and Gaines, 1932). Cavitation involves the tearing apart of a column

of fluid, and the production in it of regions of vapor or gases. The

effects of cavitation are much more evident at very high frequencies

of sound (ultrasound) then at audible frequencies.

Why sound waves do inhibit growth in some species, but may have

no effect or may even stimulate grov/th in other species of plants is

not really known. Weinberger and Measures (1970) found differences

in growth in the same species of wheat when the frequency of sound

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was varied. Below a certain frequency no inhibition of growth or

germination could be seen. Above this frequency inhibition of growth

and germination did occur. The frequency and amount of energy required

to produce these effects appears to vary widely between species.

In this experiment there is no significant interaction (Tables

11, 16, and 21; pages 23, 28, and 33). This indicates very strongly

that the observed effects, i.e. inhibition of root elongation in

cucumbers and oats, is due to sound, i.'o inhibition of root elongation

is observed for wheat. The data for cucumbers and oats, however, point

to a most dramatic inhibition of root elongation.

Germination in cucumbers, oats, ana wheat treated by 6G0 Hz sound

is not significantly effected (Tables 22, 23, and 24; pages 34, 35,

and 36). In cucumbers the data is significant at the 22 per cent level,

in oats at the 84 per cent level, and in wheat at the 18 per cent

level.

In conclusion, I have rejected the null hypothesis — Uc = Ue

(no significant difference between control and experimental runs

exists) for root elongation in cucumbers and oats. On the basis of

a statistical analysis which includes all runs, I am able to state

conclusively that the reduction in root elongation is due to sound

treatment. However, the null hypothesis cannot be rejected for wheat

since no significant inhibition of root elongation occurred. In future

experiments it would be of value to be more selective in the choice

of seeds. If only those seeds which were morphologically and genetically

similar were used (such as screened seeds; there might be less

variability between runs. Experiments should be designed which

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consider the changes (if any) in anino acid concentration between

control and sound treated seeds. A microscopic examination of seed

coats to determine differences in thickness would be of value. Finally,

it would be of interest to run either cucumbers and wheat or oats and

wheat at the same time. In doing this, a species which did show root

inhibition and one which did not show root inhibition could be run

simultaneously. The null hypothesis for the effect of 600 Hz sound

on germination — U c = Ue— is not rejected.

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l it e r a t u r e c i t e d

Aleksandrovskaya, M. M. and R. A. Chezhenkova. 1970.

Electrofiziologicheskie i morfologicheskie issledovaniya

raznykh oblastei kory bol'shikh polusharii vozdeistvii

slabym zvukom. (Electrophysiological and morphological

study of different cerebral cortical parts during weak

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5o(3):312-317.

Chambers, L. A. and Gaines N. 1932. Some effects of intense

audible sound on living organisms and cells. Jour, of

Cellular and Comparative Physiology. 1:431-469.

Ellis, George Feme. 1973* The Effect of Audible Sound on the

Germination and Root Elongation of Oats and Wheat. Master’s

Thesis.

El'Piner, I. Y. 1970. Nekavitatsionnyi ul'trazvuk i ego biologicneskoe

deistvie. (Non-Cavitational ultrasound and its biological

effect). 3io Fizika 15(3):333-343.

Frings, H., C. H. Allen, and I. Rudnick. 1948. The physical

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Gnanam, A. 1959. Activation of photosynthesis in Spirogyra by

sound waves of electric bell. Prod. Symp. Algae (New Delhi).

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Goldman, D. E. and V7. W. Lepeschkin. 1952. Injury to living cells

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Koichev, K. A. 1969. Elektronnomikroskopicheskoe issledovania

raspredeleniya natriya v kortievom orange zhivotnykh, nakhodyash

chikhsys v sostoyanii otnositel'nago pokoya i v usloviyakh

zvukovogo voxdeistviya. (Electron microscope study of sodium

distribution in the organ of Corti of animals under conditions

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cell structure. Jour, of Cellular and Comparative Physiology.

40:383-397.

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Lisenkov, A. F. 1966. Vliyanie ozvuchennoi vody na seaena drevensnykh

rastenii. (The effect of sound-treated water on seeds of woody

plants). Soviet Plant Physiology. 13(4):728-729.

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sound frequencies on the growth of Marquis spring wheat. Canadian

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of the violin on the growth of Mimosa puaica L. Proc. Indian

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of the leaves of balsam and mimosa to the musical sound of

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ultrasound frequency on the growth of synchronized cultures of

Scenedesmus obtusiuscuius. Can. Jour, of Botany. 50:361-365.

Weinberger, P. and M. Measures. 1968. The effect of two audible

sound frequencies on the germination and growth of a spring

and winter wheat. Can. Jour, of Botany. 46:1151-1162.

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