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Examining Use of Sonic Bloom Technology on the Stomata Opening of

Drought-Stressed Soybean

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DOI: 10.13005/bbra/2695

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BIOSCIENCES BIOTECHNOLOGY RESEARCH ASIA, December 2018. Vol. 15(4), p. 861-869

Examining Use of Sonic Bloom Technology on the

Stomata Opening of Drought-Stressed Soybean
Istirochah Pujiwati1, Bambang Guritno2, Nurul Aini2 and Setyawan P. Sakti3
1
Department of Agrotechnology, Faculty of Agriculture,
University of Islam Malang, 65144 East Java, Indonesia.
2
Department of Agricultural Cultivation, Faculty of Agriculture,
Brawijaya University Malang, 65145 East Java, Indonesia.
3
Physics Department, Faculty of Mathematics and Natural Sciences,
Brawijaya University Malang, 65145 East.

http://dx.doi.org/10.13005/bbra/2695

(Received: 15 November 2018; accepted: 14 December 2018)

Sonic bloom is a technology that combines high frequency sound waves and organic
nutrients, intended for better plant growth to increase itsproductivity.This study aimed to
determine the effect of sound wave frequency and drought stress on stomatal opening, nutrient
uptake efficiency through leaf, and soybean yield. We designed the research as a split plot
experiment.The main plot was sound wave frequency consisting of four levels (no frequency
imposed, frequencies 2, 4and 6 kHz. The sub-plot wasthree soil moisture contents (50,75, and
100% field capacity). We found that the interaction of frequency and soil moisture content
affected the width of stomata at the age of 30, 40 and 50 days after planting (dap), the efficiency
of nitrogen uptake, phosphorusuptake and potassiumuptake and the protein content of seeds.
The width of stomatal opening at a frequency of 4 kHz in soil moisture 100% FC showed the
highest value and was not significantly different from soil moisture 75% FC.There was a positive
correlation between exposure to plants with a frequency of 4 kHz with stomatal opening,
nutrient uptake and increased yield of soybean crops.The use of sonic bloom technology with
plant exposure at a frequency of 4 kHz could increase drought tolerance to 75% soil moisture
content. Soybean seed yield increased by 40.89% and seed protein content increased by 10.3%.

Keywords: sonic bloom, frequency, soil moisture content, nutrient uptake efficiency, soybean yield.

Soybeans are one of the important organic nutrients to make plants grow better so
agricultural commodities in Indonesia after rice as to increase their productivity. This technology
and corn. At present the consumption of soybeans utilizes natural sound waves with high frequencies
for the people of Indonesia is 2.2 million tons per that are able to stimulate the leaf mouth (stomata)
year and only 30% of domestic soybean products wide open so as to increase the rate and efficiency
while 70% are still imported. Therefore, efforts of absorption of leaf fertilizer which is beneficial
are needed to increase soybean productivity. for plants. In other words, this technology is a
Sonic bloom is a technology that combines the way to improve photosynthetic efficiency and
exposure of certainfrequency sound waves and end photosynthesis results in order to increase the

*Corresponding author E-mail: isti.wati@yahoo.co.id

This is an Open Access article licensed under a Creative Commons license: Attribution 4.0 International (CC-BY).
Published by Oriental Scientific Publishing Company © 2018
862 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018)

amount of production with good quality (Irianiet fertilization practice include: low absorption,
al, 2005; Widyawatiet al., 2011). especially in leaves whichhave thick cuticles,
One of the obstacles in soybean cultivation slipped fertilizer solution from the surface of
is the availability of water, because the soil hydropobic leaves, the solution tends to get easily
moisture was low, it resulted stunted growth, dried from the leaf surface, and leaf fertilization
high loss of flowers, and pods. In accordance cause damage to the leaves. Responses of varying
with photosynthesis, water is the main raw results in application of leaf fertilization have been
photosynthetic material in which it is needed in the widely reported. This may be related to the wrong
photolysis process, water, then, will be decomposed time of application, improper use of fertilizer
and produces electrons which are then used in ingredients, and unavailability of nutrients in the
dark reactions. Water also affects the opening of soil, as well as environmental conditions. The
the stomata. Plants that grow in low soil moisture efficiency of leaf fertilization can be influenced
conditions or drought stress,its stomata’s opening by the type of fertilizer, concentration and pH of
is relatively smaller, so CO2 absorption is low, the solution, application time, surfactant use and
consequently its photosynthesis is not optimal and compatibility with other chemical compounds
plant growth is inhibited (Khan et al., 2016). (Oosterhuis, 2009).
Water shortages in plants occurs because Dobermann (2007) provides several
there is not enough water in the media and approaches to calculate fertilizer efficiency,
excessive transpiration or a combination of these including: absorption efficiency, physiological
two factors. In the field, even though it is sufficient efficiency, and agronomic efficiency. Absorption
in the homeland, plants can experience stress efficiency is a comparison between nutrients
(lack of water). This happens if the absorption absorbed from fertilizer and the amount of fertilizer
speed is not balanced with the water-lossduring given, stated in percentagescales. Absorption
the transpiration process. In general, plants will efficiency figures are useful as a correction factor
show a certain response if they experience drought in fertilization recommendations. Physiological
stress. The response of plants to water stress is efficiency is useful for assessing plant responses
largely determined by the level of stress and the in optimizing nutrients derived from fertilizers to
phase of plants growth when they experiencing produce products. While the agronomic efficiency
stress. There are two kinds of responses that can is useful to assess how much increase in production
improve the status of water when plants experience is achieved from each amount of fertilizer added.
dry conditions, namely; (1) plants change the The efficiency of nutrient use can be expressed in
distribution of new assimilates to support root several ways. Mosier (2004) describes 4 agronomic
growth at the expense of canopy, so as to increase indices which are generally used to explain nutrient
root capacity to absorb water and inhibit leaf use efficiency, namely: partial factor productivity
expansion to reduce transpiration; (2) plants will (PFP, states kg yield for each kg of fertilizer used);
regulate the degree of opening of the stomata to agronomic efficiency (AE, kg increase in crop yield
inhibit water loss through transpiration (Brittlate, for each kg of fertilizer applied); apparentrecovery
2007). efficiency (RE, kg of fertilizer absorbed for each
Leaf fertilization practice has several kg of fertilizer given) and physiological efficiency
advantages, including low cost, fast response by (PE, kg increase in yield for each kg of fertilizer
plants, only needs a small amount of fertilizer, absorbed). The term crop removal efficiency, which
can be combined with the application of other is the nutrient percentages used to produce products
chemical compounds, and becomes very important from nutrients that are applied, is also often used
when soil conditions become a problem and (Roberts, 2008).
root growth is inadequate (Oosterhuis, 2009). Information about the use of sonic bloom
On the other hand, the leaffertilization practice technology in accordance with stomatal opening
has disadvantageswhere leaves might get burnt, at drought stress conditions has not been widely
causesolubility problems, and only a small studied. If the stomata can widely open followed
amount of nutrients can be applied at one time. by leaves fertilizationprocess, the efficiency of the
Wojcik (2004) added that some problems in leaf nutrient uptake can be further enhanced. In relation
 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018) 863

to the soybean’s stomata opening in drought stress Experimental design

conditions, it is necessary to assess the amount of The study usedSplit Plot Design, with
soil fertilizer given through sonic technology can main plot isgiven frequency of sound waves in four
be tolerated so as to produce optimal quantity and levels: F0= no frequency, F2=frequency 2 kHz, F4
quality of soybeans. =frequency 4 kHz and F6 =frequency 6 kHz. While
the sub plotconsisted of soil moisture in 3 levels:
MATERIALS AND METHODS L50 = 50% Field Capacity (FC), L75 = 75% FC, and
L100 = 100% FC. The exposure of sound waves is
Experimental place carried out from 08:00a.m. for about 20 minutes.
The study was conducted from February Soybean plants were exposed to
to June 2018 in the green house of the Faculty frequencyof sound waves from the age of 20 days
of Agriculture, University of Islam Malang, and up to 70 days after planting (dap) and treated in 10
Laboratory of Physiology, Faculty of Medicine, days interval. The exposure was carried out for 20
University of Islam Malang, Dinoyo-Lowokwaru minutes. After that,the soybean plants were sprayed
Malang, East Java province of Indonesia with with 2 g/liters of liquid fertilizer (Growmore).The
an altitude of about 540 meters above sea level. observation of the width of the stomatal opening
The research location was located at 07059’South using the molding method (replica) with fixation
Latitude and 112036' East Longitude. The average using a clear nail.The observed yield variables
temperature is 240C in the morning, 310C in the werefresh weight and dry weight of seeds, protein
afternoon and 250C in the afternoon, with humidity content of seeds. The supporting variables observed
of 57% - 83%. The average rainfall is 1.883 mm/ were nutrient uptake efficiency.
year with solar radiation intensity ranging from Statistical analysis
7.036 lux to 696.4 x 102 lux. Clay texture with The statistical analysis usedwas the
a ratio of 11% sand: 34% dust: 55% clay. NTotal analysis of variance (ANOVA) testwith split plot
0.46%, PBray 296.95 mg.kg-1 and K 1.98 me.100g-1. design, and the mean difference was tested further
Water content at pF 2 = 0.55 g.g-1 and pF 4.2 = 0.29 by using Honestly Significant Difference (HSD)
g.g-1 test at the 5% level.

Table 1. Average Stomata Width in the Interaction of Frequency and Soil

Moisture Content

Frequency (kHz) Soil Moisture Stomata Width (µm)

Content (% FC) 30 dap 40 dap 50 dap

No frequency 50 8,57 a 6,80 ab 6,50 a

75 9,73 ab 7,87 b 6,53 a
100 10,00 ab 8,00 b 6,47 a
50 8,67 a 6,17 a 6,83 ab
2 75 10,80 b 8,73 bc 8,57 b
100 11,00 bc 8,60 bc 9,53 b
50 8,67 a 5,93 a 5,40 a
4 75 12,30 bc 9,90 c 9,23 b
100 12,63 bc 10,10 c 10,03 b
50 9,20 ab 6,47 ab 6,03 a
6 75 9,43 ab 8,60 bc 9,60 b
100 9,47 ab 8,77 bc 9,67 b
HSD 5% 1,78 1,66 1,87

Notes: - The numbers accompanied by the same letter in the same column show no significant
difference in the 5% HSD (Honest Significan Difference) test -   dap: days
after planting
864 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018)

RESULTS AND DISCUSSION the lowest average width of stomatal opening for
all levels of frequency of sound wave exposure.
Stomata width Efficiency of nutrient uptake through leaves
The interaction of sound wave frequency The nutrient uptake efficiency of nitrogen,
and soil moisture content significantly affected the phosphorus and potassium was very significantly
average of stomata width at the age of 30, 40 and affected by the interaction of frequency and soil
50 days after planting (dap). moisture content.
The width of stomatal opening at 4 kHz The highest efficiency of nitrogen uptake
in soil moisture 100% FC showed the highest was achieved at 4 kHz frequency treatment at 100%
value and was not significantly different from soil soil moisture content, and it was not significantly
moisture 75% FC. In soil moisture 50% FC shows different from the 6 kHz frequency at 100% soil

Fig. 1. Efficiency of N uptake on the interaction of frequency and moisture content

Table 2. Average Efficiency of Nutrient Uptake on Interaction of

Frequency and Soil Moisture Content

Frequency Soil Moisture Efficiency of Nuptake Efficiency of Puptake Efficiency of K uptake

(kHz) Content (%FC) (%) (%) (%)

50 7,42 a 13,53 c 2,09 c
No frequency 75 10,26 a 13,13 c 2,05 c
100 28,24 d 26,52 f 4,08 f
50 9,21 a 5,54 a 0,91 a
2 75 14,18 b 9,69 b 1,56 b
100 30,70 de 25,10 ef 3,91 ef
50 21,32 c 15,04 c 2,35 c
4 75 34,69 e 22,98 e 3,56 e
100 58,46 f 45,92 h 7,05 h
50 14,51 b 4,28 a 0,75 a
6 75 30,77 de 18,70 d 2,95 d
100 59,29 f 38,98 g 6,02 g
HSD 5% 4,19 2,27 0,35

Note: The numbers accompanied by the same letter in the same column show no significant difference in the 5%
HSD test
 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018) 865

moisture content. Meanwhile, the lowest nitrogen with 100% soil moisture content. The lowest
uptake efficiency was resulted in non-frequency elemental potassium uptake at each frequency
treatment with the moisture content of 50% FCand level occurred in soil moisture content of 50% FC
it was not different fromwithout frequency of 75% (Figure 3).
FC and frequency of 2 kHz at moisture content of Fresh Weight and Dry Weight of Seeds
50% FC (Figure 1). The average fresh weight and dry weight
The highest efficiency of phosphorus of seeds was not influenced by the interaction of
uptake was obtained at 4 kHz frequency interaction frequency and soil moisture content. Separately, the
with 100% moisture content. Meanwhile at effects of both treatments are presented in Table 3.
each frequency level, 50% soil moisture content The average high fresh and dry weight
produced the lowest nutrient uptake efficiency of seeds achieved at 4 kHz was not significantly
(Figure 2). different from 6 kHz, with an increase in yields of
Potassium uptake had the same pattern as 40.89% and 30.67% respectively. While at moisture
phosphorus uptake, where the highest absorption content of 100% field capacity, yield fresh weight
efficiency was obtained at a frequency of 6 kHz and dry weight of seeds were not significantly

Fig. 2. Efficiency of P uptake on the interaction of frequency and moisture content

Fig. 3. Efficiency of K uptake on the interaction of frequency and soil moisture content
866 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018)

different from moisture content of 75% field The lowest seed protein in the treatment without
capacity with increasing yields of 35.56% and exposure to sound waves with soil moisture content
28.89% respectively. of 50% field capacity (Figure 4).
Seed Protein Content To see the relationship between the width
Seed protein content was significantly of the stomatal opening and the nutrient uptake
affected by the interaction of the frequency and efficiency given through the leaves, soybean
moisture content of the soil. The average protein growth and yield are presented in Table 5 below.
content in seeds is presented in Table 4. There was a positive correlation between
The highest seed protein content was the width of the stomatal opening and the efficiency
produced at the exposure treatment at a frequency of N, P and K nutrient uptakeand growth (which
of 6 kHz at 100% soil capacity, increasing by 10.3% is represented by the relative growth rate variable)
from the description of the variety of 41.78%. and the fresh weight and dry weight of the seeds.

Table 3. Average Fresh Weight and Dry Weight of Seeds at Frequency and
Soil Moisture Content

Treatment Fresh Weight of seeds Dry Weight of seeds

per Hectare (t.ha-1) per Hectare (t.ha-1)

Frequency (kHz)
No frequency 2,39 a 1,89 a
2 2,32 a 1,92 a
4 3,17 b 2,66 b
6 2,94 b 2,51 b
HSD 5% 0,34 0,34
Soil Moisture Content (% FC)
50 2,16 a 1,71 a
75 2,90 b 2,43 b
100 3,05 b 2,60 b
HSD 5% 0,39 0,30

Note: The numbers accompanied by the same letter in the same column show no significant
difference in the 5% HSD test (Anjasmoro variety production potential: 2.25-2.30 t.ha-1)

Fig. 4. Seed protein content in the interaction of frequency and soil moisture content
 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018) 867

Discussions the water will move into the guard cell and the
turgidity of the guard cell increases, so the stomata
In general, at all frequency levels, at 100% open. Although the drought conditions spurred
soil moisture content, the field capacity resulted the closure of the stomata, in the condition of soil
in a wide opening of the stomata which was not moisture 75% the field capacity of stomata guard
significantly different from the soil moisture cells can still maintain their turgidity. The results
content of 75% field capacity. This shows that the of research by Budisantoso and Proklamasiningsih
turgidity of guard cells can still be maintained at a (2003) concluded that the use of sonic bloom
decrease in soil moisture content by 25%. Misraet technology can eliminate the effects of drought to
al., (2015) suggested that stomatal opening and 75% of the soil moisture content available.
closing is determined by increasing and decreasing The interaction between the frequency
turgor pressure in guard cells in response to and level of soil moisture significantly affected the
environmental conditions and biological systems. efficiency of fertilizer absorption given through the
Marschner (2012) suggested that potassium ions (K leaves and the protein content of seeds. The highest
+) along with other anions such as Ca2 +, H + have efficiency of elemental absorption of nitrogen,
a very important role in regulating the pressure of phosphorus and potassium at all frequency levels
guard cells (guard cells) during stomatal opening. at 100% soil moisture content, was not significantly
Increasing the concentration of K + in the guard cell different from that produced at 75% soil moisture
causes the osmotic potential of the cell to decrease, content. This related to the optimal opening of
stomata to the level of soil moisture content of 75%
field capacity, thus it also supportedthe optimal
Table 4. Average Soybean Seed Protein Content in the
Interaction of Frequency and Soil Moisture Content nutrient uptake through stomata. The results of the
correlation analysis showed a positive relationship
between the width of the stomatal opening and
Seed Protein Content (%) the nutrient uptake efficiency, growth and yield of
Frequency (kHz) Soil Moisture Content soybean plants. Wide opening stomata supports
50% 75% 100% the entry of nutrients given through the leaves
(Oosterhuis, 2009) so as to produce photosynthetic
No Frequency 35,53 a 36,23 b 36,50 b products which are reflected in the increase in plant
2 43,00 c 43,70 d 44,47 de
growth and yield. The yield component which
4 43,70 d 44,57 de 45,23 de
6 43,97 d 45,00 de 46,07 f was influenced by the interaction between the
HSD 5% 0,69 frequency and level of soil moisture was the seed
protein content, where the highest seed protein was
Note: The numbers accompanied by the same letter in the same obtained at a frequency treatment of 6 kHz with
column show no significant difference in the 5% HSD test soil moisture content of 100% field capacity which

Table 5. Correlation Coefficients between Stomata Width, Nutrient Uptake Efficiency,

Relative Growth Rate (RGR), Fresh Weight of Seeds (FWS) and Dry Weight of Seeds (DWS)

Stomata Uptake Uptake Uptake RGR FWS DWS

Width of N of P of K

Stomata Width 1.000** 0.574* 0.618* 0.616* 0.611** 0.775** 0.124
Uptake of N 0.574* 1.000** 0.937** 0.930** 0.024 0.877** 0.900**
Uptake of P 0.618* 0.937** 1.000** 0.974** 0.013 0.818** 0.827**
Uptake of K 0.616* 0.930** 0.974** 1.000** 0.019 0.813** 0.822**
RGR 0.611* 0.024 0.013 0.019 1.000** 0.013 0.015
FWS 0.775** 0.877** 0.818** 0.813** 0.013 1.000** 0.991**
DWS 0.124 0.900** 0.827** 0.822** 0.015 0.991** 1.000**

Note: ** very significantly different at p <0.01, significantly different at p <0.05

868 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018)

was 46.07%. At a frequency of 4 kHz the protein 35.56% from its potential production of 2.25 t/ha.
content of seeds produced on soil moisture content This result was not significantly different from
of 100% field capacity was 45.23% not significantly soil moisture treatment 75% field capacity with a
different from soil moisture 75% field capacity of production increase of 28.89%. This is in line with
44.57%. At a frequency of 2 kHz, the highest seed the result of research conducted by Budisantoso
protein content was produced in soil moisture and Proklamasiningsih (2003) where the use of
content of 100% field capacity of 44.47% while in sonic bloom technology can eliminate the effects
the treatment without frequency the protein content of drought to 75% soil moisture content available.
of seeds on soil moisture content of 100% field Soil moisture is an environmental factor that limits
capacity was only 36.50%, and it was not different plant growth and yield. The response of plants to
from 75% field capacity which was 36.23%. There water shortages can be identified from its metabolic
was an increasing content in soybean protein up activity, morphology, and physiology. Chlorophyll
to 10.3% in plants which were given sound waves content of the plant showed the lowest yield in soil
exposure compared to the its varieties description moisture treatment 50% field capacity. The results
of 41.78 - 42.05%. Chowdhuryet al. (2014) of the research by Song Ai and Banyo (2011) stated
explained that there is a very close relationship that the decreasing chlorophyll content is one of the
between sound waves and plant growth. Sound plant’s physiological responses to water shortages.
waves with certain frequencies have a significant The decreasing chlorophyll content relates to
effect on biological, biochemical, physiological photosynthetic device activity, and it decreases
and gene expression activities in plants. Further plant photosynthesis rate. Ashraf and Harris (2013)
Yiyaoet al. (2002) explained that the division and added that chlorophyll as a photosynthetic pigment
growth of plant tissue is strongly influenced by would be damaged in the presence of drought stress
dissolved proteins in the tissues. Accumulation resulting in a decrease in the efficiency of light
of dissolved protein content affects cell division, absorption in both photosystem-I and photosystem-
enzyme content and level of metabolism. Zhao II, so the photosynthetic capacity is reduced. Light
et al. (2003) added that sound waves at specific energy is absorbed by chlorophyll and converted to
frequencies increase dissolved protein and sugar chlorophyll fluorescence, despite the fact that the
content in the Dendranthemamorifolium callus level of chlorophyll fluorescence is not more than
cytoplasm. The optimum frequency of sound 1-2% of the total absorbed light.
waves stimulates an increase in soluble protein
content and callus growth in Chrysanthemum calli CONCLUSION
(Bochuet al., 2001; Yiyaoet al., 2002; Yi et al.,
2003). Sound waves exposure at frequency of 5 Sonic bloom technology was able to
kHz for 4 weeks increases the amount of alanine increase the wide opening tolerance of soybean
and glycine in the grain endosperm tissue compared stomata to drought stress conditions of 75% field
to control plants (Measures and Weinberger, 1973). capacity. Nutrient uptake efficiency increased to
Separately the frequency of sound waves 52.4% for nitrogen, 31.9% for phosphorus, and
of 4 kHz with 6 kHz produced insignificant fresh 32.2% for potassium than without using sonic
weight and dry weight. This is in line with the bloom. The increasingnutrient uptake efficiency led
research conducted by Martens et al. (1982)that to the increasing seed production to 40.89% and
only a certain frequency range could stimulate plant soybean seed protein content increased to 10.3%.
growth and yield. Furthermore Chowdhuryet al.
(2014) added that sound with a certain frequency ACKNOWLEDGMENT
and intensity that has a positive influence on plant
biological activities such as seed germination, root I received Doctoral Research Grant
extension, plant height, callus growth, enzyme and from the Directorate Research and Community
hormone activity and gene expression. Meanwhile Service, Ministry of Research, Technology and
the soil moisture content treatment of 100% field Higher Education of Indonesia, contract numberis
capacity increased the fresh weight of the seed by 211/B.07/U.V/LPPM/2018.
 Pujiwati et al., Biosci., Biotech. Res. Asia, Vol. 15(4), 861-869 (2018) 869

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