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Cinnamon powder: an in vitro and in vivo evaluation
of antifungal and plant growth promoting activity
Jolanta Kowalska &Józef Tyburski &
Joanna Krzymińska &Magdalena Jakubowska
Accepted: 18 October 2019
#The Author(s) 2019
Abstract In this study, the effects of organic powder of
Cinnamomum zeylanicum on the development of Botry-
tis cinerea and its influence on tomato plants were
evaluated. The cinnamon bark powder and its water
suspensions and filtrates were used at 0.5 and 1% rates.
After 6 days of the start of an in vitro experiment the
mycelium growth was inhibited by both 0.5 and 1%
cinnamon water filtrates - to a greater degree in the case
of the higher concentration, by 54.4 and 81.4%, respec-
tively. Spraying with cinnamon water filtrates positively
influenced the growth of plants both in the greenhouse
and the field. Antifungal activity of cinnamon was
proved in the greenhouse tests - the disease symptoms
of grey mould on infected tomato plants decreased. The
fresh weight of non-inoculated tomato plants treated
with cinnamon filtrates was significantly higher than
control plants (17.17 g compared to 12.83 g) showing
a stimulating effect of cinnamon filtrates. In the case of
inoculated plants due to treatment, their weight in-
creased from 7.83 to 10.50 g. In the field experiment,
tomato plants sprayed six times with cinnamon were
better developed than the control plants. The most sig-
nificant effect was observed for Hamlet variety - the
mean number of leaves was higher by 27.3% and the
mean number of branches by 19.7% compared to the
untreated control plants. Thus it was proved that cinna-
mon powder has potential to inhibit B. cinerea growth
and also has a stimulating effect for tomato plants.
Keywords Botrytis cinerea .Basic substance .
Mycelium growth inhibition .Grey mould symptoms
inhibition .Tomato vegetative development
Introduction
There is a worldwide trend to explore new alternatives
to control plant diseases, giving priority to methods that
reduce disease symptoms and avoid negative side ef-
fects on human health resulting from excessive applica-
tion of synthetic fungicides. The use of food substances,
which are also classified as basic substances, demon-
strating a protective action in growing crops, is an
alternative method for organic farmers or sustainable
agriculture farms.
The possibility of limiting the development of path-
ogenic fungi by applying agents, which are natural and
alternative to pesticides, is a popular research direction.
But also the use of plants or plant products as fungicides
has been of a great importance for some time and needs
more attention (Zaker 2016). New pesticides, including
natural product-based pesticides are being discovered
and developed to replace the compounds lost due to
the new registration requirements (Regulation (EC)
No1107/2009 of the European Parliament and of the
Council of 21 October 2009 concerning the placing of
Eur J Plant Pathol
https://doi.org/10.1007/s10658-019-01882-0
J. Kowalska :J. Krzymińska (*):M. Jakubowska
Institute of Plant Protection National Research Institute, 20
Władysława Węgorka Street, 60-318 Poznań,Poland
e-mail: j.krzyminska@iorpib.poznan.pl
J. Tyburski
University of Warmia and Mazury in Olsztyn, 2 Michała
Oczapowskiego Street, 10-719 Olsztyn, Poland
plant protection products on the market and repealing
council directives 79/117/EEC and 91/414/EE’2009).
The use of natural products is one of biological control
methods in fungal disease control. The method is still
not common and investigations are required to find
suitable plants that can be used to control pathogenic
fungi (El-Mougy Nehal and Abdel-Kader Mokhtar
2007; Kowalska 2011; Remlein-Starosta et al. 2016).
Antifungal activity of essential oils (EOs) such as
cinnamon, marigold, basil and spearmint was assessed
against pathogens such as Botrytis cinerea Pers., Fusar-
ium gramineraum Corda,FusariumoxysporumSchltdl.
and Sclerotinia sclerotiorum (Lib.) de Bary (Kocic-
Tanackov et al. 2011; Al-Taisan et al. 2014; Ghalem
2016). The use of EOs and various plant extracts is
aligned with current thinking on the future of agriculture
in the EU, and with consumer preference for natural
products (Brenes and Roura 2010). The antimicrobial
properties of EOs have been widely reported (Dorman
and Deans 2000; Mourey and Canillac 2002;Rotaetal.
2004). Besides antibacterial properties, EOs or their
components have exhibited anti-parasitic (Pandey et al.
2000;Pessoaetal.2002), and insecticidal
(Konstantopoulou et al. 1992; Karpouhtsis et al. 1998)
properties. EOs are widely used in various fields, in-
cluding the cosmetics, pharmaceutical, and food indus-
tries (Harkat-Madouri et al. 2015). The constituents of
these compounds have antibacterial, viricidal, fungicid-
al, anti-parasitic, insecticidal, and medicinal properties
which are considered to be plants protectants (Bakkali
et al. 2008).
Cinnamon oils and extracts showed good antifungal
properties against important plant diseases. Wilson et al.
(1997) found that among 49 essential oils tested,
Cinnamomum zeylanicum J. Presl demonstrated a great
antifungal activity against B. cinerea.Cinnamomum
zeylanicum commonly known as cinnamon,refers to a
tropical evergreen tree as well as a bark that is extracted
from the plant. Substances such as cinnamon have been
known to the mankind for a very long time. They are
universally recognized as harmless and safe for human
consumption. Cinnamon powder is considered to have
high potential as a biological control agent (Sikes et al.
2005; Xing et al. 2010; Kocić-Tanackov and Dimić
2013).
B. cinerea Pers. Fr. (teleomorph Botryotinia
fuckeliana (de Bary) Whetzel) is an economically im-
portant phytopathogenic fungus and a cause for a broad
spectrum of plants diseases (van Kan 2006; Bolton et al.
2006;Deanetal.2012). It can cause serious damage in
over 200 plant species both in pre- and post-harvest
stages and is difficult to control due to various modes
of attack and many hosts (Williamson et al. 2007). Very
few reports concerning the use of commercially avail-
able powdered cinnamon can be found in the literature.
Therefore, the aim of the study was to evaluate the
potential of commercially available cinnamon powder
originating from organic crops used in the liquid form in
different concentrations as a control agent against
B. cinerea andgrowthstimulatorinbothinvitroand
in vivo conditions.
Materials and methods
Plant pathogenic fungus preparation
For all tests B. cinerea strain 2235 (plant source: tomato
stem) from the Bank of Plant Pathogens in the Institute
of Plant Protection, Poland, collection was used. Pure
cultures were maintained on Potato Dextrose Agar,
Sigma-Aldrich (PDA) media at 4 °C and sub-cultured
at monthly intervals. Ten days old cultures were used for
the in vitro experiment.
For the greenhouse and field experiments conidia
were harvested from ten days old cultures. The concen-
tration was determined using the Thoma slide and ad-
justed to 3.1 × 10
5
ml
−1
. The resulting suspension was
used to inoculate the plants.
Cinnamon suspensions and filtrates preparation
The preparation method was developed by the authors.
Commercially available organic cinnamon
(C. zeylanicum) powder (manufactured by Dary Natury,
Grodzisk Poland and certified by Ekogwarancja PTRE
sp. z o.o.) was used to obtain water suspensions and
filtrates. For the in vitro experiment modified PDA
media was used. The cinnamon suspensions were pre-
pared using warm sterile distilled water (26 °C) and
added to PDA media to obtain 0.5 and 1% cinnamon
PDA suspensions (0.5 and 1 g of cinnamon added to
20 ml of water and 80 ml of the PDA media, respec-
tively). Later cinnamon water suspensions were filtered
through a nylon flour mesh (150 μm) and added to PDA
to obtain 0.5 and 1% cinnamon filtrates PDA suspen-
sions (0.5 and 1 g of cinnamon powder added to 20 ml
of water, filtrated; 20 ml of the filtrate added to 80 ml of
Eur J Plant Pathol
the PDA media). For the greenhouse and field experi-
ments 0.5% water filtrate was used. Five grams of
cinnamon was added to 1 l of warm tap water (26 °C)
and filtered through a flour mesh. The filtrate was pre-
pared shortly before spraying. Water suspensions and
filtrates were chosen due to their practical application
mode. The filtrates are clearer and do not cause prob-
lems with the foliar spraying using a sprayer with a
nozzle, while in the case of a suspension containing
powder particles, it is necessary to strain the suspension
twice before using a fine mesh and to apply with a
coarse nozzle.
In vitro biological control activity evaluation
The evaluation of biocontrol activity of cinnamon water
suspensions and filtrates against the growth of
B. cinerea mycelium was done (El-Mougy Nehal et al.
2004). Disks (0.5 cm diameter each) from pure cultures
of the pathogenic fungus were placed in the middle of a
90 mm Petri dish containing PDA with or without the
addition of cinnamon water suspensions and filtrates in
five replications. Fungi were grown at 23 °C. Mean
colony diameter (mm) was measured after 3 and 6 days
(i.e. until a control Petri dish was fully covered by the
mycelium; the maximum growth was 85 mm –the
diameter of the dish excluding the diameter of the initial
culture).
Plant development evaluation in greenhouse conditions
Experiments were performed in pots according to
methods developed by the authors. Ten tomato plants
(Ozarowska variety) in the 3–4 leaf phase were planted
into each pot. Each repetition consisted of five pots.
Altogether five repetitions were made. Later, plants
were inoculated by foliar spray with a water suspension
of B. cinerea spores (3.1 × 10
5
ml
−1
), 1 ml per plant.
Three and six days after inoculation water filtrates of
cinnamon at 0.5% was sprayed on the plants surface.
The control plants were: a) in the first control group,
inoculated with B. cinerea and sprayed with water; and
b) in the second control group, not inoculated and
sprayed with cinnamon water filtrates. After an addi-
tional 5 days, the foliage lesions were assessed (the
presence of B. cinerea was verified microscopically).
Theimpactofsprayingwithcinnamononplantswas
evaluated as the above ground fresh weight of plants
collected after 3 and 6 weeks of cultivation.
Plant development evaluation in field conditions
The field trial was carried out on a certified organic farm
according to methods developed by the authors. Three
field tomato varieties (Agro, Country and Hamlet) were
used. Seedlings were planted into soil in June, at the 5-
leaf phase, with five plants from each cultivar used in
each repetition. Altogether four repetitions were made
for each cultivar. Six sprayings of water cinnamon fil-
trates (0.5%) were performed at 4–5 day intervals. The
controls were not treated with cinnamon filtrates. The
number of leaves and branches was assessed after one
and three months of planting to evaluate plants
development.
Statistical analysis
A one-way analysis of variance (ANOVA) was used to
test significance with Tukey-Kramer’s post hoc test for
multiple comparisons. A pvalue <0.05 was accepted as
statistically significant.
Results and discussion
In the laboratory experiment, the potential of cinnamon
water suspensions vs. cinnamon filtrates in reducing
B. cinerea growth on PDA medium was evaluated.
Three days after PDA inoculation the highest inhibition
effect of mycelial growth was obtained while using 1%
of cinnamon water suspensions and 1% of cinnamon
filtrates, which reduced the fungus lesion by 64 and
47.8%, respectively (Table 1). Taking into account all
the treatments (with water suspension and filtrate and
the two concentrations –0.5 and 1%), prolongation of
the experiment from 3 to 6 days resulted in an average
increase of the mycelium lesions diameter from 10.8 to
60.3 mm. In the same time the changes in the effective-
ness of the two cinnamon formulations were observed.
After 6 days the effect of cinnamon suspensions (both
0.5 and 1%) was very weak to none - in fact the statis-
tical analysis proved no curative effect. The positive
effect was found using cinnamon filtrates –0.5% filtrate
concentration decreased B. cinerea mycelium growth by
54.4% and 1% filtrate gave a lesions reduction by 81.4%
(Table 1). In other words, the best inhibitory effect in an
in vitro study was obtained while using 1% filtrate of
cinnamon powder.
Eur J Plant Pathol
In the greenhouse conditions the disease symptoms
decreased on the plants treated with cinnamon water
filtrates compared to the control plants which were
inoculated and sprayed only with water. On average
1.75% of the plants treated with cinnamon were infected
compared to 2.28% of control plants. Additionally,
spraying with cinnamon water filtrates influenced posi-
tively the growth of plants. After six weeks of the
application the fresh weight of plants treated with cin-
namon was significantly higher than the control. Non-
inoculated plants treated with cinnamon weighed on
average 17.17 g compared to 12.83 g for the non-
inoculated control treated with water (33.8% increase
of mass). Plants inoculated with B. cinerea and treated
with cinnamon weighed on average 10.50 g compared
to 7.83 g for the inoculated control treated with water
(34.1% increase of plant weight) (Table 2).
In the field experiment the number of leaves on tomato
plants counted one month after planting showed a differ-
ent pattern of development in the grown varieties. Variety
Agro decreased number of leaves by 10.3% showing an
inhibitory effect, while varieties Country and Hamlet
increased number of leaves by 5.2 and 27.3%, respec-
tively, showing stimulating effect (Table 3). After two
months of growth the number of branches on the plants
was counted. The varieties Agro and Hamlet showed a
positive reaction to cinnamon water suspension spraying
and increased number of branches by 15.3 and 19.7%,
respectively. In the same time variety Country did not
respondtocinnamonspraying–the cinnamon spraying
did not affect the number of branches. The vegetative
growth response of tomato plants to cinnamon water
suspension spraying depended on the phase of plants
development and variety.
According to other studies, both in vitro and in
planta, essential oils derived from cinnamon proved to
have bio-control properties against many pathogens
(Siripornvisal et al. 2009; Hyldgaard et al. 2012).
Shabana et al. (2015) tested cinnamon essential oil
against B. cinerea in vitro and noted a significant sup-
pression of the pathogen growth up to 7 days, compa-
rable to the chemical fungicide Topsin M70-WP. Allam
et al. (2017) noted that higher concentration of cinna-
mon completely inhibited the mycelial growth of
Tabl e 1 The influence of cinnamon water suspensions and filtrates on an average lesion diameter [mm] of B. cinerea measured on day 3 and
6 after beginning of in vitro tests
Treatment (cinnamon formulation) 3 days after beginning of test 6 days after beginning of test
Lesion diameter, mm Reduction of lesion, % Lesion diameter, mm Reduction of lesion, %
Control 16.10 ± 1.29a 0.0 85.00 ± 0.00a 0.0
0.5% water suspension 10.10 ± 1.43a 37.3 76.90 ± 18.11a 9.5
1% water suspension 5.80 ± 0.84b 64.0 85.00 ± 0.00a 0.0
0.5% water filtrate 13.70 ± 3.63a 14.9 38.80 ± 8.06b 54.4
1% water filtrate 8.40 ± 0.65b 47.8 15.80 ± 2.02c 81.4
Averaged data 10.8 40.9 60.3 36.3
Values in each column followed by the same letter are not statistically different P=0.05
Data are presented as mean ± standard deviation (SD)
Tabl e 2 The average fresh weight (g) of tomato plant inoculated with B. cinerea and treated with 0.5% cinnamon water filtrates in the pot
experiment
Number of weeks of tomato
cultivation in pots
Control
(water spraying)
Non-inoculated plants sprayed
with cinnamon filtrate
Inoculated plants
sprayed with tap water
Inoculated plants sprayed
with cinnamon filtrate
Tomato fresh weight after
3 weeks,
2,14 ± 0.17a 2,09 ± 0.11a 1,71 ±0.14a 1,85 ± 0.14a
Tomato fresh weight after
6 weeks
12.83 ± 0.87b 17.17 ± 1.07a 7.83 ± 0.62c 10.50 ± 1.12b
Values in each row followed by the same letter are not statistically different P=0.05
Data are presented as mean ± standard deviation (SD)
Eur J Plant Pathol
B. cinerea in vitro. The antifungal potential of cinnamon
was confirmed for others pathogens by other authors
(Barrera-Necha et al. 2009;Xingetal.2014;Sarkhosh
et al. 2018). The significant antifungal activity of cinna-
mon oil (both in vitro and in vivo) against various
Fusarium species which was proportional to its concen-
tration was shown. In the study by Horváth et al. (2013)
the cinnamon oil effectively inhibited mycelial growth
of Fusarium head blight of winter wheat in vitro. Jiang
et al. (2013), Al-Taisan et al. (2014) and Moraes et al.
(2018) examined the inhibitory effects of cinnamon
against S. sclerotiorum among other essential oils and
microelements in in vitro experiments and as soil
application. All concentrations of cinnamon
completely inhibited the mycelial growth of the
fungus. According to Moraes et al. (2018) the result
was comparable to a commercial fungicide
(thiophanate-methyl +chlorothalonil). Ojaghian et al.
(2014) proved that crude extracts of cinnamon are able
to reduce in vitro mycelial growth, sclerotial
myceliogenic and carpogenic germination of
S. sclerotiorum. In other studies, during the liquid bio-
assay,C.zeylanicumwas fungicidal against pathogens
isolated from banana, including Colletotrichum musae
(Berk. & M.A. Curtis), Lasiodiplodia thebromae (Pat.)
Griffon & Maubl., and Fusarium proliferatum
(Matsush.) Nirenberg ex Gerlach & Nirenberg
(Ranasinghe et al. 2002); exerted antifungal activity
towards Oidium murrayae Hosag.,U.Braun&
Rabindran (Chu et al. 2006) and inhibited conidial ger-
mination of Colletotrichum gloesporioides Penz.) Penz.
& Sacc. (Barrera-Necha et al. 2008). In in vitro exper-
iments it was found to have a good mycelial inhibition
of the corn rot F. oxysporum f.sp. gladioli (Massey)
(Barrera-Necha et al. 2009) and to be highly effective
against the growth of Rhizoctonia solani Kühn (Nguyen
et al. 2009), as well as to have excellent antifungal
activity against early blight of tomato Alternaria solani
Sorauer (Yeole et al. 2014). The investigations of Wang
et al. (2014) showed that cinnamon microemulsions had
a high in vivo control activity against gray mould of
pears B. cinerea. The results obtained by the authors
also confirmed activity of cinnamon towards control of
grey mould in all experimental areas. In presented re-
search water filtrates of cinnamon were more efficient
than water suspensions, this fact has two implications, i)
it is more applicable to use and, ii) water filtrates should
be used in future tests including also chemical analysis
of compounds. So far in planta studies on cinnamon as a
bio-control agent against B. cinerea mainly reported its
effectiveness on post-harvest products. It applied to such
plants as pears (Wang et al. 2014), citrus fruit (Bouchra
et al. 2003), peppers and tomatoes (Kong et al. 2016). In
most cases cinnamon oil was used. It had no significant
negative effect on plant product qualities such as firm-
ness and colour. The control for decay incidence and
lesion diameter was reported to be promising.
While existing literature focuses on antifungal prop-
erties of cinnamon, the authors have noticed a signifi-
cant positive effect of cinnamon filtrate spraying on
plant branching in the field experiment. In addition to
the fact that the antifungal, antibacterial and pest deter-
ring properties of cinnamon improved the general well-
being of plants and thus enabled improved growth and
branching, there might be another process undergoing
on a different level. Therefore, further studies are needed
to determine the cause.
To conclude, in laboratory conditions cinnamon wa-
ter filtrates were more effective against B. cinerea than
water suspensions and based on literature it seems to be
as effective as cinnamon oil. Further studies are required
to recognise the mechanisms of cinnamon improving
plants growth and development. Cinnamon water fil-
trates have also proven to be effective in in planta
experiments, including both greenhouse and field toma-
toes, where the stimulating effect on plant development
Tabl e 3 The influence of cinnamon water filtrate (0.5%) on vegetative development of tomato plants after 1 month (July) and 3 months
(September) of application under field conditions
Combination Control (water spraying) Cinnamon water suspension spraying
Variety Agro Country Hamlet Agro Country Hamlet
Number of leaves 12.6 ± 0.58a 11.6 ± 0.08b 11.0 ± 0.71b 11.3 ± 0.55b 12.2 ± 0.50a 14.0 ± 0.71a
Number of branches 14.3 ± 0.58 17.8 ± 1.62a 12.7 ± 1.10b 16.5 ± 0.55a 18.0 ± 1.65a 15.2 ± 0.82a
Values in each row followed by the same letter are not statistically different P=0.05
Data are presented as mean ± standard deviation (SD)
Eur J Plant Pathol
and plant health was confirmed. Formulation of water
suspensions and its water filtrates were chosen because
of its practical application of being used by growers,
while the use of cinnamon oil can be difficult in practise.
The inhibitory effect of cinnamon water filtrates may be
used for practical application provided the treatments
are carried out several times at 4–5 day intervals. This
frequency of treatments is sufficient to maintain the
inhibitory effect on the developing pathogen. This con-
clusion has practical implications especially for organic
growers who have less plant protection products to use
than the farmers growing crops the conventional way.
Compliance with ethical standards
Conflict of interest The authors declare that they have no con-
flict of interest.
Research involving human participants and/or animals N/A
Informed consent N/A
Open Access This article is distributed under the terms of the
Creative Commons Attribution 4.0 International License (http://
creativecommons.org/licenses/by/4.0/), which permits unrestrict-
ed use, distribution, and reproduction in any medium, provided
you give appropriate credit to the original author(s) and the source,
provide a link to the Creative Commons license, and indicate if
changes were made.
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