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INTERNATIONAL JOURNAL OF AGRICULTURE & BIOLOGY
ISSN Print: 1560–8530; ISSN Online: 1814–9596
15–583/2016/18–3–515–520
DOI: 10.17957/IJAB/15.0117
http://www.fspublishers.org
Full Length Article
To cite this paper: Demirkaya, M., W. Stumpf and K. Jezik, 2016. Influence of organic fertilizers on physiological quality and rooting of snapdragons
(Antirrhinum majus). Int. J. Agric. Biol., 18: 515‒520
Influence of Organic Fertilizers on Physiological Quality and Rooting of
Snapdragons (Antirrhinum majus)
Mustafa Demirkaya1*, Werner Stumpf2 and Karoline Jezik2
1Department of Agriculture, Safiye Cikrikcioglu Vocational College, Erciyes University, 38039, Kayseri, Turkey
2Department of Crop Sciences, Division of Vegetables and Ornamentals, University of Natural Resources and Life Sciences,
Gregor-Mendel-Strasse 33, 1180, Vienna, Austria
*For correspondence: mustafad@erciyes.edu.tr
Abstract
In this study, the effects of organic fertilizers on internal quality and rooting of snapdragons (Antirrhinum majus) were
investigated. Seedlings of ‘White Rocket’, ‘Rocket Gold’, ‘Orchid’, ‘Red Stone’ and ‘Red Rocket’ species were transplanted
into 12 L pots filled with 1:1 sand: soil mixture at the beginning of March 2011. Two different liquid biological fertilizers
were used. The first one is the Effective Microorganisms (EM®) consisting of lactic acid bacteria (Lactobacillus casei and
Lactobacillus plantarum), photosynthesis bacteria (Rhodopseudomonas palustris) and fermentation bacteria (Saccharomyces
cervisiae); the second one is Rhizo Vital 42® consisting of Bacillus amyloliquefaciens bacteria. The physiological quality of
the petals was investigated. With regard to petal P-dissipation values, while EM treatments yielded almost identical results
with the control treatment, Rhizo Vital 42 treatments had lower yield values than the control treatment. Both organic fertilizer
treatments improved rooting and effects of EM treatments on rooting were found to be significant. Biophoton measurements
were also performed over the petals and photoluminescence values of fertilizer treatments of all species were found to be
lower than the control plants. In other words, organic fertilizer treatments improved the quality by decreasing the stress levels
in petals. © 2016 Friends Science Publishers
Keywords: Dissipation value; Biophoton emission; Effective microorganisms; Rhizo Vital 42
Introduction
There are different microorganisms exist like bacteria and
fungi from which normally developing plants are not
affected. However, there are also some bacteria and fungi
that can cause diseases. Moreover, there are different
species of bacteria and fungi that stimulate plant
development and health (Jezik et al., 2011).
Since intensive agricultural practices may result in
various environmental problems, organic farming practices
are getting more and more popular nowadays. Thus,
biofertilizers are commonly recommended instead of
synthetic fertilizers (Karakurt et al., 2011). Plant growth
promoting rhizobacteria (Bacillus species) are also
commonly used to promote plant growth and development
(Priest et al., 1987).
Effective Microorganisms (EM)
The EM of the present study is a commercial organic
fertilizer containing regenerative microorganisms.
Microorganisms enable a suitable environment for
fermentation and play an important role in plant quality and
soil productivity. Fermentative fragmentation is stimulated
by the cease of decomposition. That means edible
microorganisms exist on the soil, optimum results can be
extracted, diseases can be prevented and high quality
products can be produced. Over all, EM generally works in
anaerobic areas in which the problems such as
decomposing, putrefaction etc. appear. Thus, they can be
used in anaerobic areas more efficiently. EM can be used to
revive the soil, increase soil fertility and remove odor,
increase the soil temperature, clean water etc.
Microorganisms can be used in all areas of life. When EM
have no other tasks, it is not difficult for pathogens to
disappear. It is also easy to remove EM from the soil
through negative signals (Lorch, 2010).
EM consists of lactic acid bacteria (Lactobacillus casei
and Lactobacillus plantarum), photosynthesis bacteria
(Rhodopseudomonas palustris) and fermentation bacteria
(Saccharomyces cervisiae). EM do not include any
genetically modified organisms. Following the preparation,
it must be used in a week (Jezik et al., 2011).
Rhizo Vital 42 – Bacillus Amyloliquefaciens FZB 42
These bacteria exist in close vicinity of the roots. There are
several types of these microorganisms and they are called
Demirkaya et al. / Int. J. Agric. Biol., Vol. 18, No. 3, 2016
516
rhizosphere microorganisms. The best known example is a
fungus called Mycorrhiza. Another kind is Bacillus
amyloliquefaciens (Kilian and Raupach, 1999). Bacillus
amyloliquefaciens Rhizo Vital 42, which has antimicrobial
activity, can produce a secondary change by high capacity
and it strengthens the plant as a gram-positive bacterium
(Chen et al., 2007). The plant root-colonizing-strain Bacillus
amyloliquefaciens FZB 42 is an environmental strain which
is distinguished from the domesticated model organism
Bacillus subtilis 168 by its ability to stimulate plant growth
and to suppress plant pathogenic organisms (Idriss et al.,
2002). FZB 42 genome analysis revealed the presence of
numerous gene clusters involved in the synthesis of non-
ribosomal synthesized cyclic lipopeptides (Koumoutsi et al.,
2004) and polyketides (Chen et al., 2006; Schneider et al.,
2007) with distinguished antimicrobial action.
The present study was conducted to investigate the
effects of Rhizo vital 42 and effective microorganism
treatments on internal quality of petals and rooting of 5
different snapdragon varieties.
Materials and Methods
Experimental Details and Treatments
Experimental material: Experiments were carried out in
Jedlersdorf in the gardens of the University of Natural
Resources and Life Sciences. Jedlersdorf, which lies at a sea
level of 162 m, has a precipitation of around 550 mm/year
and an average temperature of 9.8°C. The sun shines hours
are 1800 hours/year. The climate is dry, warm and windy
with a yearly average of 3.2 m/s (Jezik et al., 2010). The
experiments consisted of 12 plots and each plot consisted of
5 rows (varieties) with 6 pots (replications). Four plots were
treated with EM, 4 plots with Rhizo Vital 42and 4 plots
were used for monitoring as control without any treatments.
Treatments: To reduce the impacts of the surroundings, a
boundary area of snapdragons (A. majus) was set up. To
suppress weeds, thick black mulching was laid out on the
ground of whole experimental area. At the beginning of
March 2011, 5 different varieties of snapdragon seeds were
sown in viols. The varieties were ‘White Rocket’ (white),
‘Rocket Gold’ (yellow), ‘Orchid’ (purple), ‘Red Stone’
(light-red) and ‘Red Rocket’ (dark-red). Germination took 2
weeks at a temperature of 15 – 20°C. At the end of March
2011, the seedlings were transplanted to 12 L pots filled
with a compost-sand mixture in the ratio of 1:1 at beginning
of May 2011. Organic fertilizers (EM and Rhizo Vital 42)
were applied to the plant pots at the following doses:
initially a 10 L pitcher was half-filled with tap water, then
10 ml EM was collected with a syringe, injected into the
pitcher and stirred with a wooden stick. Water was added to
fill the pitcher completely to the 10 L mark. Similar
implementations were performed for Rhizo Vital 42, but
this time 40 ml Rhizo vital 42 was injected into 5 L of water
and then completed to 10 L. The prepared mixtures were
applied weekly to each pot in 500 mL doses. Since the
plants were smaller at the beginning of the experiment, the
initial dose was arranged as 250 mL per plant for the first
three treatments (Jezik et al., 2011). Specified doses of EM
and Rhizo Vital 42 were applied weekly until the plant dry
out date of 24th August 2011. As soon as the plants started
blooming, all the closed buds were harvested at a size of
about 2 cm, all the stamina were removed and the plain
petals were stored in a deep freezer at -21°C. Harvesting
was also stopped on 24th August due to dry out of the plants.
The effects of Rhizo vital 42 and EM treatments on
rooting were investigated as well. Plants were cut from the
soil surface and the pots were emptied at the end of August
2011. A scale from 1 to 6 was formed to evaluate the root
development (Table 1). In this way, the effects of treatments
on root development were evaluated.
The root data were statistically analyzed by ANOVA
and correlation analyses. SPSS 13.0 for Windows and LSD
tests were also used for a mean separation.
Then the frozen petals were put in the refrigerator for a
day for defrosting. After that, the photon measurements
were made. For the measurements of the dissipation values,
extracts had to be made with a juice separator (Multi-press
automatic MP 80, BRAUN, Germany). After centrifuging
and filtering, the extracts were ready for the measurements.
Photoluminescence by Single Photon Counting
From a biophysical point of view, electromagnetic
interaction is dominant in biological systems and is
described as thenon-equilibrium thermodynamic
organization of biochemical substance (Feynman, 1985).
Therefore, living systems emit and absorb electromagnetic
fields called photons. This emission of the light fields of
biosystems is known as “biophoton emission“, sometimes
called as “ultraweak bioluminescence”.
Photoluminescence measurements were performed in
4 replications by using a single photon counting device
(Photomultiplier Hamamatsu R 943-02, high voltage
Tennelec TC 952, amplifier-discriminator HamamatsuC
3866, measuring card Tennelec Nucleus MCS II) developed
at the Atomic Institute of the Technical University in
Vienna Geissler (1999) now at the University of Natural
Resources and Life Sciences, Vienna.
Measuring glasses were filled with 6 g of defrosted
petals, which were radiated with the light of a quicksilver-
high-pressure-lamp (HOL-R delux, 80 watt, OSRAM,
Germany) at a distance of 35 cm for 90 s. Then the samples
were put into the air-tight measuring chamber of the photon
measuring arrangement. All this was executed under a rapid
timetable. The photoluminescence of a sample was
measured for 500 s in 4 replications. The integral photon
emission with its special deviation kinematics was
calculated and drawn by the software Origin Pro 8.0. Finally
the amount of integral photon emission was interpreted with
regard to internal quality: the lower the amount, the higher
Influence of Organic Fertilizers on Snapdragons (Antirrhinum majus). / Int. J. Agric. Biol., Vol. 18, No. 3, 2016
517
the quality.
In order to quantify different deviation kinematics, the
declining curves were fitted by the double exponential
function
(y = A1.exp (-x/t1) +A2.exp (-x/t2)). (1)
Dissipation Value
The dissipation value is used to interpret the energetic
conditions of plants and can be calculated from pH-value,
electrical conductivity and redox potential (Velimirov,
2003). Therefore, the dissipation value is usually interpreted
as follows: the lower it is the higher the energy potential and
the more suitable it is for human nutrition (Zimmermann,
2003). In other words, smaller dissipation values indicate
better qualities (Hoffmann, 2005). Only 10 mL petal
solution (extract) could be extracted in the present study.
However, about 30 – 40 mL extract is required for the
measurements. Therefore 20 mL distilled water was added
to complete the extract volume to 30 mL. Then the
electrochemical measurements of these sample solutions
were performed by using the pH-Meter multi 340i from the
WTW Company (Germany). The instrument features three
electrodes, one to measure the redox potential and
temperature (SenTixORP, WTW, Germany), one to
measure the pH value and the temperature (SenTix 41,
WTW, Germany) and one to measure the electrical
conductivity (TetraCon 325, WTW, Germany). The pH
electrode should initially be calibrated with the calibration
solutions for pH values of 4.01 and 7 (by placing into buffer
solutions with these pH values) and the conductivity
electrode should be calibrated by placing in to 0.01 mol HCl
(hydrochloric acid). Before and after the calibrations, the
electrodes were washed with distilled water and dried with
fine paper towels. Then a magnetic stirrer was placed into
each solution sample and placed over a magnetic field.
Redox potential, pH value, electrical conductivity and
temperature were measured simultaneously. The entire
measurement data (180 values) were transferred into Excel.
Then the dissipation values were calculated by using the
following equation. The averages of the first and the last
value of each parameter (pH-value, redox potential,
electrical conductivity and temperature) were used to
calculate the dissipation value. The electrical conductivity
values were converted from μS/cm to mS/cm. Then
following equations was used:
Dissipation value [µW] =Eh2/Ω
Eh = measured redox potential + electrodeconstant +
temperature correction factor.
Temperature correction factor = (measured
temperature -25) * (-0.71) Ω= 1 /(R *0.001)
rH value=Eh/constant+ 2 * pH (2)
Where: rH = redox potential based on the pH-value
Ω= electrical resistance
Electrode constant= 207 Constant =29.07 Eh =redox
potential [mV] – relative to the potential of the normal
hydrogen electrode R= electrical conductivity [mS/cm]
E=redox potential [mV].
Results
Integral Photon Emission of Photoluminescence
Compared to control treatment, EM treatments reduced
photon emission by 31.25% in ‘Red Rocket’, by 19.23% in
‘Red Stone’, by 16.33% in ‘Orchid’, 13.00% in ‘White
Rocket’ and finally by 53.00% in ‘Rocket Gold’. Rhizo vital
42 treatments decreased photon emissions by 68.75% in
‘Red Rocket’, 76.92% in ‘Red Stone’, 73.33% in ‘Orchid’,
77.22% in ‘White Rocket’ and 69.35% in ‘Rocket Gold’
variety. Such findings revealed that Rhizo vital 42
treatments improved petal quality much more than EM
treatments (Fig. 1).
Dissipation Value
Through the application of EM and Rhizo Vital 42, positive
results could be achieved in the varieties ‘Orchid’ and ‘Red
Rocket’. The dissipation values of the petals were
significantly lower than those of the control. In this case, it
is considered that these two varieties have a high energy
potential and therefore have a high quality. In the varieties
‘Golden’ and ‘Red Stone’, only the Rhizo Vital 42
treatments could decrease the dissipation value of the petals
and hence achieve a better quality. White Rocket could not
be positively influenced by any of the treatments. On the
contrary, the dissipation values of the petals were
significantly higher than those of the control (Table 2).
Compared to the control, EM and Rhizo Vital 42
treatments caused significantly lower dissipation values in
the leaves of all varieties, except ‘White Rocket’. This
represents a lower energy potential and consequently a
lower quality for ‘White Rocket’ and the opposite for the
other varieties. The most positive impacts of EM treatments
on leaves were observed in ‘Red Stone’ with a value of 570
and most positive impacts of Rhizo vital 42 treatments were
observed in ‘Orchid’ with a value of 554. The results
revealed that Rhizo vital 42 has the most positive effects on
the petals and leaves of ‘Orchid’ (Table 3).
Effects of Rhizo Vital 42 and Effective Microorganisms
on Rooting
EM treatments had more positive impacts on root
development than Rhizo Vital 42. The highest increase
in rooting was found in ‘White Rocket’ treated with EM.
However ‘Orchid’ treated with EM and ‘White Rocket’
treated with Rhizo vital 42 also showed a high increase.
Also in ‘Red Rocket’ and ‘Golden’ treated with EM, the
increase was high. Rhizo Vital 42 could not significantly
increase the root development of ‘Red Rocket’ and
‘Golden’. In ‘Red Stone’, neither of the two treatments
could achieve better rooting (Table 4).
Demirkaya et al. / Int. J. Agric. Biol., Vol. 18, No. 3, 2016
518
Discussion
Popp (1991) indicated that biophoton measurements could
be used to assess bioenergetics status and tissues of plants.
The researchers also indicated that the lower the biophoton
emission, the higher the internal quality. Since Rhizo vital
42 and EM treatments decreased biophoton emissions, they
improved internal quality (Fig. 1). Zimmerman (2003)
indicated that the lower the p-dissipation value, the higher
the energy potential will be thus more suitable for human
nutrition. In another study, Hoffmann (2005) indicated
lower P-dissipation values as the indicator of better quality.
Compared to control treatment, Rhizo vital 42 and EM
treatments generally decreased P- dissipation values of the
petals and leaves (Tables 2 and 3). Such findings revealed
the positive impacts of Rhizo Vital 42 and EM treatments
on internal quality parameters.
The impacts of Rhizo vital 42 and EM treatments on
root development were also investigated in this study and
the effects of EM treatments were found to be significant.
Bajwa et al. (1999) reported that EM treatments
increased the effects of vesicular arbuscular mycorrhizal
fungi (VAM) colonization in chick pea (Cicer
arietinum). Increased phosphorus and other nutrient
uptakes through VAM colonization were also reported
by other researchers (Graham and Menge, 1982; Smith
et al., 1992). Kafkas and Ortaş (2009) indicated
increased phosphorus uptakes of Pistacia species with
VAM treatments. Various researchers indicated that
phosphorus uptake improved the rooting (Lynch and
Brown, 2001; Walk et al., 2006). Compared to control
and Rhizo Vital 42 treatments, improved rooting with
EM treatments of the present study supports the
findings of previous studies. There is a certain level of
microbial activity in soil, which is necessary for proper
plant growth. Beside beneficial microorganisms,
however, harmful microorganisms are also present in soils.
Beneficial microorganisms inoculated into soils in order to
remove the harmful ones from the soil and consequently
reduce the stresses exerted on plants, increase internal
quality and positively affect root development. The
present study clearly indicated the positive impacts of
Rhizo Vital 42 and EM treatments on plant root
development and internal quality parameters. However,
such positive impacts strictly depend on plant species,
regional climate conditions, soil conditions,
implementation doses and timing.
Jezik et al. (2011) carried out a research on lettuce
with organic fertilizers and observed 10% yield increase. It
was concluded in this study that Rhizo Vital 42 and
Effective Microorganisms could be used effectively as
organic fertilizers in ornamental plant production. They may
have positive impacts on inner quality parameters but the
effects may vary based on species, varieties and
treatments. Therefore, further research is needed on
implementation doses and timing. Further research is
also recommended to assess the impacts of such organic
fertilizers on common vegetables. Since EM and Rhizo
Vital 42 treatments do not have any proven negative
impacts on soil and human health, further studies with
other plants like vegetables should be conducted for
sustainable agriculture.
Acknowledgments
The authors acknowledge YOK (The Council of Higher
Table 1: Scale for evaluating effects of Rhizo Vital 42 and
Effective Microorganisms on root development
Scale number
Meaning
0
no rooting
1
little rooting
2
slightly more rooting
3
halfway rooted through
4
lot of rooting
5
very significant rooting
6
completely rooted through
Table 2: Effects of treatments on dissipation of petals
Dissipation values [µW]
Variety
Control
EM
Rhizo Vital
'Red Rocket' (dark-red)
426
344
380
'Red Stone' (light-red)
442
464
417
'Orchid' (purple)
502
443
373
'White Rocket' (white)
442
535
527
'Golden' (yellow)
513
588
462
Table 3: Effects of treatments on dissipation of leaves
Dissipation values [µW]
Variety
Control
EM
Rhizo Vital
'Red Rocket' (dark-red)
890
728
725
'Red Stone' (light-red)
850
570
669
'Orchid' (purple)
747
675
554
'White Rocket' (white)
610
621
697
'Golden' (yellow)
655
587
571
Table 4: Effects of Rhizo Vital 42 and Effective Microorganisms on rooting
Varieties
Treatment
'Red Rocket' (dark-red)
'Red Stone' (light-red)
'Orchid' (purple)
'White Rocket' (white)
'Golden' (yellow)
Control
1.79±1.23abc
1.33±0.57cd
1.17±0.89d
1.58±1.16 bcd
1.38±0.53cd
EM
2.14±0.82a
1.14± 0.48d
1.61±0.48bcd
2.13±0.84 a
1.76±0.57abc
Rhizo Vital 42
1.83±0.96abc
1.21±0.43d
1.29±0.82d
2.04±0.97 ab
1.50±0.83cd
Mean ± standart deviation; Values indicated with different letters are significantly different (P < 0.05)
Influence of Organic Fertilizers on Snapdragons (Antirrhinum majus). / Int. J. Agric. Biol., Vol. 18, No. 3, 2016
519
Education), for the scholarship provided to M. Demirkaya
and to Prof. Dr. Herbert Klima and DI. Dr. Alexander
Geissler for explaining the photon measuring arrangements.
The research activities were conducted at the University of
Natural Resources and Applied Life Sciences, Vienna.
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