Gesunde Pflanzen

https://doi.org/10.1007/s10343-020-00534-2

ORIGINAL ARTICLE / ORIGINALBEITRAG

Effect of Microwave Radiation on Bacteria, Fungi and Some Growth

Characteristics of Cowpea Vigna unguiculata L.
Wael Mohammed Mahdi1 · Khaled Saeed Lateef Al-Badri2 · Mustafa R. M. Alqaisi3

Received: 5 June 2020 / Accepted: 13 November 2020

© Springer-Verlag GmbH Deutschland, ein Teil von Springer Nature 2020

Abstract
In this study, an experiment was conducted to describe the effects of using microwaves and chemical fertilization (isolated
and combined) on the number of bacteria and fungi in the soil and on the growth of cowpea plants Vigna unguiculata L.
Several times of exposure to microwave radiation (0, 25, 30, 35, 40, 45 s) and several concentrations of chemical fertilizer
NPK (0, 0.9, 1.8 g 3 kg–1 soil) were carried out. After individual and combined treatments, experiments were carried out to
quantify microorganisms (bacteria and fungi). In addition, 45 days after planting, analyzes were made of the cowpea plants
for their development under different experimental conditions. The experiment was conducted in plastic pots to find out
some characteristics of plant growth. The results indicated that the microwave sterilization resulted in the complete killing
of bacteria and fungi and the fertilizer additive (NPK) to the soil had a critical role in increasing the values significantly
for the number of root nodes and the remaining plant growth characteristics measured in this study.

Keywords Bacteria · Cowpea · Fungi · Microwave sterilization · NPK fertilizer

Wirkung von Mikrowellenstrahlung auf Bakterien, Pilze und einige Wachstumsmerkmale der
Augenbohne Vigna unguiculata L.

Zusammenfassung
In dieser Studie wurde ein Experiment durchgeführt, um die Auswirkungen von Mikrowellen und chemischer Düngung
(isoliert und kombiniert) auf die Anzahl von Bakterien und Pilzen im Boden und auf das Wachstum der Augenbohnen-
Pflanzen Vigna unguiculata L. zu bestimmen. Es wurden mehrere Expositionen mit Mikrowellenstrahlung (0, 25, 30, 35,
40, 45 s) und mehrere Konzentrationen des chemischen Düngers NPK (0, 0,9, 1,8 g 3 kg–1 Boden) getestet. Nach indivi-
duellen und kombinierten Behandlungen wurden Experimente zur Quantifizierung von Mikroorganismen (Bakterien und
Pilze) durchgeführt. Darüber hinaus wurden 45 Tage nach der Pflanzung Analysen der Augenbohnen-Pflanzen bezüg-
lich ihrer Entwicklung unter verschiedenen Versuchsbedingungen durchgeführt. Das Experiment wurde in Plastiktöpfen
durchgeführt, um einige Merkmale des Pflanzenwachstums zu untersuchen. Die Ergebnisse deuteten darauf hin, dass die
Mikrowellensterilisation zur vollständigen Abtötung von Bakterien und Pilzen führte und dass der Düngerzusatz (NPK)
zum Boden eine entscheidende Rolle bei der signifikanten Erhöhung der in dieser Studie gemessenen Werte für die Anzahl
der Wurzelknoten und die übrigen Pflanzenwachstumsmerkmale spielte.

Schlüsselwörter Bakterien · Augenbohne · Pilze · Mikrowellensterilisation · NPK-Dünger

 Mustafa R. M. Alqaisi
dr.mustafa.r.m@tu.edu.iq
2
Department of Physics, College of Education, University of
1
Department of Biology, College of Education, University of Samarra, Salah-AL-Din, Iraq
Samarra, Salah-AL-Din, Iraq 3
Department of Horticulture, College of Agriculture Tikrit
University, Salah-AL-Din, Iraq

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Introduction
Cowpea (Vigna unguiculata L.) production suffers from
extreme despair due to pests and diseases. The main prob-
lems are caused by Striga gesneriads parasitic weeds;
Macrophomina phaseolina (Tassi) Goid, a fungal disease,
and bacterial disease caused by Xanthomonas campestris
pv. vignicola (Pammel) Dowson, with various viral dis-
eases. Data indicate that M. phaseolina attacks are more
significant when the cowpea plant is already weakened
by high temperature and severe drought conditions. The
fungi will be present for long periods in the soil and on
the seeds, which is essential for the epidemiology of the
disease. Various control methods, such as pesticides and
breeding, are used to combat these diseases (Adam 1990;
Iqbal and Mukhtar 2020).
Soil is a complex environment because it contains many
living and non-living components. Especially beneficial and
harmful microorganisms (Erabi and Ali 2014), and bush
plants compete for cultivated crops (Sahin 2014). So, re-
searchers in the field of soil turn to find different methods
of sterilization of the soil contribute to the elimination of
various pathogens. Including bacterial and fungal, which
lead to significant losses during the growth stages of plants
(AL-Samarrai et al. 1988; Erabi and Ali 2014), or when
experimenting with the effect of bio fertilization technique
to study the effect of the organism used.
Sterilization of the soil is the process of eliminat-
ing worms, nematodes, fungi, bacteria, and other plant
pathogens (Nutter 1957). There are various methods of
soil sterilization, some of which have a selective effect
on soil type, but not others. Such as fungicides, which
should be considered when using collateral damage such
as toxicity to humans and residual impact. As well as high
cost and, effort to achieve (Almeida et al. 1994), or the
use of solar energy (Blacksmith et al. 2016) and others
may cause general annihilation of organisms such as the
use of an autoclave, gama rays, UV rays, microwave rays,
formalin, methyl bromide, vapour, etc. Komarova et al.
(2008) studied the effect of soil sterilization in different
ways on microbial content. Erabi and Ali (2014) found that
the results of the sterilization methods and their numbers
had a significant impact on the numbers of bacteria and
fungi, and the characteristics of the soybean plant. The
number of bacteria and fungi decreased before and after
planting by sterilization with formaldehyde and formalin
compared to unsterilized soil. The results also showed that
sterilization by two methods led to a significant decrease
in germination percentage, plant height, dry weight of
plants and total roots compared to the non-sterile treat-
ment. Public awareness about the adverse impacts of bush
pesticides and harmful microorganisms and the possible
adverse effects of their residues on the environment have
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W. M. Mahdi et al.
encouraged researchers to make numerous attempts to de-
velop microwave-based technology (MW) in large-scale
seed and soil treatment. It has revolutionized agriculture
(Nelson 1996).
The microwave is part of the electromagnetic radiation
spectrum. The frequency ranges from 300 MHz to 300 GHz
with a wavelength of 1 m to 1 mm (Halmagyi et al. 2017).
This technetium has many uses such as industry and wire-
less communications Laboratory also besides it uses in ex-
periments and biosciences (Chen 2006). It has other uses,
mainly to sterilize laboratory instruments (Tisserat et al.
1992), as well as soil sterilization of pests and pathogens,
as well as their very rapid drying (Diprose 2001). In the field
of crop production, microwaves are applied using individual
devices that allow weeding in field conditions (Velázquez-
Martí et al. 2008) and sterilization in greenhouses to pre-
vent the occurrence and transmission of fungal and viral
infections (Soriano-Martin et al. 2006; Oza et al. 2008).
Treatments with this technique are practically applicable
in disinfecting soil and seeds of vital pollutants in addition
to their role in reducing weeds that may accompany plant
growth, as well as being inexpensive financially (Brodie
et al. 2007). Heat source, temperature, and duration of ex-
posure can affect the physical and chemical properties of
chemical fertilizers (Song et al. 2012), and various soil ele-
ments (Ferriss 1984; Gibson et al. 1988; Hendricks and Pas-
coe 1988; Zhang and Shen 2006). Microwave soil releases
some absorbed nutrients and dramatically increases the ef-
ficiency of converting organic nitrogen to metal (Zieliński
and Zieliński 2010). Khan et al. (2016) in their study of
soil exposure to microwave radiation (2.45 GHz) that the
sterilization gave a 175% increase in dry biomass and 96%
in grain yield compared to untreated soil.
This study aims to investigate the effects of microwave
in soil sterilization, and the vital nutrients added NPK for
the impact of this technique on the number of root nodes
and growth of cowpea plant grown in plastic pots.
Material and Methods
Soil Sterilization Based on Microwave Oven
A field experiment was conducted using plastic pots with
a capacity of 3 kg per pot. It was filled with mixed soil
sifted with a 2 mm sieve after taking a sample to deter-
mine its physical, chemical and biological characteristics
as in Table 1. The soil was fertilized with NPK compound
fertilizer at two levels (0.9 and 1.8 g 3 kg–1) in addition to
non-fertilization. Then five seeds/pot of local cowpea seeds
(black-eyed pea cultivar) were planted at a depth of 2 cm
according to the method described from De Wilde et al.
(2017). The pots planted for sterilization in the microwave
Effect of Microwave Radiation on Bacteria, Fungi and Some Growth Characteristics of Cowpea Vigna unguiculata L.

Table 1 Physical, chemical, and biological characteristics of the soil Design and Analysis of the Experiment
Characters Unit Value
EC Ds.m–1 2.28 An experiment was designed according to the randomized
pH – 7.75 complete block design (RCBD) with three replicates. The
Organic matter g kg–1 15.54 units of tests were: first factor exposure times 0, 25, 30, 35,
Available N mg kg –1 26.35 40 and 45 s and second factor addition of chemical fertil-
Available P 9.03 izer NPK at different levels 0, 0.9, 1.8 g 3 Kg–1 soil. The
Available K 88.5 experiment results were analyzed based on the Duncan test
Sand g kg –1 51.6 at a probability level of 0.05.
Silt 19.0
Clay 29.4
Soil texture Loam Results and Discussion
Total number of fungi CFU g–1 6 × 10 3

Total number of bacteria CFU g–1 120 × 106 Effect of Sterilization Using Microwave Radiation on
Bacteria and Fungi in the Planted Soil of Cowpea

oven manufactured by Kenwood (1000 watts and 2400 MHz
frequency) were presented with different exposure times: 0,
25, 30, 35, 40 and 45 s in addition to the control treatment.
Then, all pots were removed from the oven and given
equal amounts of plain water as long as the plants remain
in the pot. Dilution and dish counting method was used to
calculate total bacterial count using nutrient agar medium
and full fungi preparation using the potato agar dextrose
medium in soil according to the method described by Ishaq
and Khan (2011). Plant data collection of growth parameter
like the number of root nodes (node plant–1), plant height
(cm plant–1), dry vegetable and root weight (g plant–1) were
calculated after 45 days of planting (Motaleb et al. 2020).
The results of Fig. 1 show a significant decrease in the num-
ber of bacteria and fungi in the soil due to sterilization at
100% energy level. The 40 s time sterilization treatment
gave a significant reduction in the microbial population
of 0.02 × 105, 0.01 × 107 CFU g–1 dry soil compared to the
unsterile treatment that gave 46 × 105 and 6 × 107CFU g–1
dry soil for bacteria and fungi, respectively. The results
of Fig. 1 show that microwave sterilization has killed bacte-
ria and fungi completely, especially in time 45 s. The effect
of microwave radiation on microbial soil totals is associ-
ated with many unknown factors (Nelson 1996). Microwave
propagation in the soil is influenced by many factors such as
soil moisture content, total density, organic matter content
and temperature (Nelson 1996; O’neill and Jackson 1990).

Fig. 1 Effect of sterilization

using microwave radiation in the
planted soil of cowpea

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 W. M. Mahdi et al.

Table 2 Effect of microwaves Time of Mean

Chemical fertilizer
sterilization and fertilizer NPK exposure
on number of root nodes of NPK NPK NPK
[seconds] 0.0 g 3 kg–1 soil 0.9 g 3 kg–1 soil 1.8 g 3 kg–1 soil
cowpea plant (node plant–1)
0 1.00ef 1.00 ef 1.33 def 1.11 b
25 2.66 bce 6.00 a 4.00 abc 4.22 a
30 1.66 def 1.00ef 1.33 def 1.33 b
35 4.33 ab 3.33 bcd 2.33 bcdef 3.33 a
40 0.33 f 1.33 def 2.00 cdef 1.22 b
45 1.33 def 2.33 bcdf 2.00 cdef 1.88 b
Mean 1.88 a 2.50 a 2.16 a –

Table 3 Effect of microwave Time of Mean

Chemical fertilizer
sterilization and chemical fertil- exposure
izer NPK on height of cowpea NPK NPK NPK
[seconds] 0.0 g 3 kg–1 soil 0.9 g 3 kg–1 soil 1.8 g 3 kg–1 soil
after 45 days of planting (cm
plant–1) 0 13.00 d 13.33 d 12.66 d 13.00 c
25 20.83 abc 24.16 ab 16.33 cd 20.44 b
30 20.00 bc 22.33 ab 20.00 bc 20.77 ab
35 23.83 ab 23.83 ab 22.50 ab 23.38 a
40 20.90 abc 23.65 ab 25.50 a 23.35 a
45 20.16 bc 22.66 ab 23.50 ab 22.11 ab
Mean 19.78 b 21.66 a 20.08 ab –

The effect of a microwave on the growth of microorgan-
isms depends primarily on the frequency of the wave and
the total energy absorbed by the microorganism (absorbed
dose). Also, besides when the microwave is used in high
frequency and high energy for a long time, the thermal ef-
fect probably is the effecter on killed the bacterial cells or
yeasts. Rahi and Rich (2008) and Brodie et al. (2015) have
shown that microwave soil treatment reduces or eliminates
weed seed, nematode, soil bacteria and pathogenic fungi
such as Fusarium oxysporum and Sclerotium rolfsii.
Effect of Microwave Sterilization and NPK Fertilizer
on the Number of Root Nodes of Cowpea
Table 2 shows the effect of different sterilization times and
NPK fertilizer and their interaction on the number of root
nodules of cowpea after 45 days of transplantation. The re-
sults showed significant differences between exposure times
25 and 35 s compared with other times and control treat-
ment. The exposure time 25 and 35 s give a higher mean of
several root nodes which was 4.22 and 3.33 nodes plant–1,
respectively, compared with non-sterile control treatment
which gave the lowest average number of node reached to
1.11 nodes plant–1.
As for chemical fertilization, there were no significant
differences between fertilizer and non-fertilizer treatments.
The overlap between the sterilization and fertilization
process also showed significant differences in some cases.
Where sterile treatment with 25 s exposure and fertilized at
0.9 g 3 kg–1 soil gave the highest average number of nodes
of 6.00 nodes plant–1 compared to sterile treatment with 40 s
exposure and non-fertilizer which gave the lowest average.
The number of nodes was 0.33 nodes plant–1. The steril-
ization of the soil significantly on this indicator, its role
may be, removing pathogens and some harmful organisms
and competition for plant growth (Hess et al. 2018; Brodie
et al. 2015), as shown in Table 1. As for the interfering
effect between microwave sterilization and fertilization in
addition to the role of sterilization, in other fertilizer stud-
ies, the application of phosphorous fertilizers has consider-
ably improved the number of nodules (Nkaa et al. 2014).
These observations are actual since phosphorus initiates the
formation of nodules and influences the efficiency of rhizo-
bium-leguminous symbiosis, thus improving nitrogen fixa-
tion (Haruna and Aliyu 2011). Whereas, these results are
inconsistent with the findings by Abayomi et al. (2008).
Effect of Microwave Sterilization and Chemical
Fertilizer NPK on Cowpea Plant Height
Table 3 shows the effect of varying sterilization times, NPK
fertilization and their interaction on cowpea height 45 days
post-cultivation. The results showed that there were signif-
icant differences between the different exposure time and
control treatment. As the exposure time of 35 s was the
highest mean higher by 23.38 cm plant–1 compared to the
non-sterile comparison treatment, which gave the lowest
mean height of 13.00 cm. plant–1. This may be attributed

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Effect of Microwave Radiation on Bacteria, Fungi and Some Growth Characteristics of Cowpea Vigna unguiculata L.

to the killing of harmful organisms in soil and bushes that
compete with cowpea. In addition to increased accumula-
tion of nitrogen and other nutrients in the soil as a result of
the thermal effect of microwave (Khan et al. 2016), Cooper
and Brodie (2009), Wainwright et al. (1980), Cawse and
Crawford (1967) who demonstrated that microwave steril-
ization leads to increased ammonium ions and sulfate ox-
idation in the soil, which may lead to improved seedlings
and improved growth.
As for chemical fertilization, the results showed sig-
nificant differences between fertilized and non-fertilized
treatments. Fertilized treatment at level 0.9 g 3 kg–1 gave
the highest average height of 21.66 cm. plant–1 compared
to non-fertilized treatment that gave the lowest average
height of 19.78 cm plant–1. These results are consistent with
Kwon et al. (2019) application of typical compound fertil-
izer (NPK) increased the plant height, This may be due to
the contribution of these elements (NPK) to the essential
components of growth factors, such as DNA.
As for the overlap between sterilization and fertilization
process, there were significant differences in many cases.
The sterile treatment with 40 s exposure time and fertilizer
level 1.8 g 3 kg–1 soil gave the highest average height of
25.50 cm plant–1 compared to the non-sterile and non-fertil-
ized control treatment, which gave the lowest average. The
height reached 13.00 cm plant–1.
Effect of Microwave Sterilization and Chemical
Fertilizer NPK on the Dry Vegetable Weight of
Cowpea Plants
Table 4 shows the impact of different sterilization times
and NPK fertilization and the interaction between them on
the dry vegetable weight. The results showed significant
differences between the different exposure time and control
treatment, the exposure time 30 sec. give the highest dry
vegetable weight, which was 1.22 g plant–1 compared with
non-sterile control treatment, which gave the lowest average
height of 0.60 g plant–1.
As for the chemical fertilization, the results showed sig-
nificant differences between fertilizer treatments level 1.8 g
3 kg–1 soil, which gave the highest average dry vegetable
weight of 1.11 g.plant–1 compared to non-fertilizer treat-
ment that offered the lowest average dry vegetable weight
of 0.88 g plant–1.
As for the interaction between the sterilization and fertil-
ization process, there were significant differences in many
cases. The sterile treatment with 25 sec. exposure time and
fertilizer level 1.8 g 3 kg–1 soil gave the highest average dry
vegetable weight of 1.40 g plant–1 compared to the non-
sterile control treatment fertilized at a 1.8 g 3 kg–1 soil level.
It gave the lowest average dry vegetable weight of 0.56 g
plant–1. These effects coincide with those obtained by Pok-
luda et al. (2018), who claimed that adding NPK increases
some of the growth characteristics, such as the dry weight

Table 4 Effect of microwave Time of Mean

Chemical fertilizer
sterilization and chemical fertil- exposure
izer NPK on the dry vegetable NPK NPK NPK
[seconds] 0.0 g 3 kg–1 soil 0.9 g 3 kg–1 soil 1.8 g 3 kg–1 soil
weight of cowpea (g plant–1)
0 0.57 de 0.67 cde 0.56 e 0.60 c
25 0.82 bcd 1.12 abc 1.40 a 1.11 ab
30 1.28 ab 1.21 ab 1.18 abc 1.22 a
35 0.93 abcde 0.93 abcde 1.17 abc 1.01 ab
40 1.01 abcde 1.15 abc 1.29 ab 1.15 ab
45 0.69 cde 0.91 abcde 1.07 abcd 0.89 b
Mean 0.88 b 1.00 ab 1.11 a –

Table 5 Effect of microwave Time of Chemical fertilizer Mean

sterilization and chemical fertil- exposure
izer NPK on the dry root weight NPK NPK NPK
[seconds] 0.0 g 3 kg–1 soil 0.9 g 3 kg–1 soil 1.8 g 3 kg–1 soil
of cowpea (g plant–1)
0 0.20 cd 0.20 cd 0.24 c 0.21 c
25 0.36 b 0.46 a 0.38 ab 0.40 a
30 0.22 cd 0.25 c 0.14 d 0.20 c
35 0.46 a 0.40 ab 0.44 a 0.43 a
40 0.44 a 0.26 c 0.22 cd 0.30 b
45 0.25 c 0.24 c 0.35 b 0.28 b
Mean 0.32 a 0.30 a 0.29 a –

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of the leaves. This may be due to the presence of nitrogen,
which has a role in stimulating photosynthesis activity.
Effect of Microwave Sterilization and NPK on Dry
Root Weight of Cowpea Plants
Table 5 shows the effect of different sterilization times and
NPK fertilization and their interaction on dry root weight of
cowpea plant after 45 days of planting. The results showed
significant differences between the different exposure time
comparative to control treatment, the exposure time 35 sec.
gave the highest dry root weight 0.43 g plant–1 compared
with non-sterile control treatment which gave the lowest
average dry root weight of 0.21 g plant–1.
As for chemical fertilization, there were no significant
differences between fertilized treatments compared to non-
fertilized treatment.
As for the overlap between the sterilization process and
fertilization, there were significant differences in many
cases. The exposure time of 35 s and non-fertilized highest
mean dry root weight of 0.46 g plant–1 followed by sterile
treatment. At the same time and fertilized at the level of
1.8 g 3 kg–1 soil, which gave an average weight root was
0.44 g plant–1 while sterile and fertilized treatment at 1.8 g
3 kg–1 soil gave the lowest average dry root weight of 0.14 g
plant–1.
This result is consistent with the results observed by
Krasilnikoff et al. (2003), Nyoki et al. (2013) This could
be attributed to the fact that phosphorus is required in
large quantities at the tips of the shoots and roots where
metabolism is high. Cell division is rapid (Ndakidemi and
Dakora 2007). Therefore, an indication that cowpea vari-
eties have used phosphorus fertilizers judiciously applied
in the growth and development processes. In addition to
that, Astuti et al. (2020) found that fertilization with NPK
increased the dry matter in the plant.
Conclusion
From this study, we can conclude that the growth traits such
as root nodes, plant height and dry weight of both plant and
plant root were greatly enhanced. The results indicated that
the microwave sterilization resulted in the complete killing
of bacteria and fungi and the fertilizer additive (NPK) to the
soil had a critical role in increasing the values significantly
for the number of root nodes and the remaining plant growth
characteristics measured in this study.
Acknowledgements The authors are thankful to the Department of
Biology and Physics, College of Education, University of Samarra,
Salah-AL-Din, Iraq to work in the laboratory and department facilities
and would like to thank the anonymous reviewers for their insightful
suggestions.
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W. M. Mahdi et al.
Conflict of interest W.M. Mahdi, K.S.L. Al-Badri and M.R.M. Alqaisi
declare that they have no competing interests.
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