Received: 6 October 2020 | Revised: 20 December 2020 | Accepted: 31 December 2020

DOI: 10.1111/jph.12979

ORIGINAL ARTICLE

The role of Cinnamomum zeylanicum essential oil,

(E)-­cinnamaldehyde and (E)-­cinnamaldehyde oxime in the
control of Meloidogyne incognita

Aline Ferreira Barros1 | Vicente Paulo Campos1 | Letícia Lopes de Paula1 |

1 1 2
Luma Alaís Pedroso | Fabíola de Jesus Silva | Júlio Carlos Pereira da Silva |
3
Denilson Ferreira de Oliveira | Geraldo Humberto Silva4

1
Department of Plant Pathology, Federal
University of Lavras (UFLA), Lavras-­MG,
Brazil
2
Department of Plant Pathology, Federal
University of Santa Maria (UFSM), Santa
Maria-­RS, Brazil
3
Department of Chemistry, Federal
University of Lavras (UFLA), Lavras-­MG,
Brazil
4
Institute of Exact Science, Universidade
Federal de Viçosa -­Campus de Rio
Paranaíba, Rio Paranaíba, Brasil
Correspondence
Aline Ferreira Barros, Department of Plant
Pathology, Federal University of Lavras
(UFLA), 37200-­0 00, Lavras-­MG, Brazil.
Email: afbarros2004@yahoo.com.br
Funding information
Coordenação de Aperfeiçoamento de
Pessoal de Nível Superior; Conselho
Nacional de Desenvolvimento Científico
e Tecnológico; Fundação de Amparo à
Pesquisa do Estado de Minas Gerais
Abstract
The search for natural nematicides that are biodegradable with little or no human
toxicity has intensified in recent years. In this context, the use of essential oils has
the potential to function as an alternative plant–­parasitic nematode control strat-
egy, and their characterization may identify new nematicidal molecules. In this study,
the nematicidal activity of Cinnamomum zeylanicum essential oil, its most abundant
biochemical component and its analogue were evaluated against Meloidogyne incog-
nita. Mean LC50 and LC95 values for C. zeylanicum oil were 49 µg/ml and 131 µg/
ml, respectively. When J2 placed in C. zeylanicum oil at its LC50 concentration were
used to inoculate tomato plants, the reduction in numbers of galls and eggs versus
samples inoculated using J2 and no C. zeylanicum oil was 98%. C. zeylanicum essential
oil reduced levels of M. incognita J2 that hatched 38%–­54%, while carbofuran (posi-
tive control) did not prevent hatching. C. zeylanicum oil applied at a concentration
of 800 µg/ml to a potted substrate containing infested tomato plants significantly
reduced numbers of M. incognita galls. The cinnamaldehyde molecule within C. zey-
lanicum oil had LC50 and LC95 values of 64 and 768 µg/ml, respectively, while LC50
and LC95 values for cinnamaldehyde oxime were 323 and 529 µg/ml, respectively.
Both cinnamaldehyde and its oxime inhibited hatching in M. incognita J2. These find-
ings indicate that C. zeylanicum essential oil, its major biochemical component, cin-
namaldehyde and cinnamaldehyde oxime (a cinnamaldehyde analogue) can be used
to reduce levels of M. incognita.

KEYWORDS

cinnamaldehyde oxime, cinnamon, nematicide, root-­knot nematodes

1 | I NTRO D U C TI O N
Worldwide losses caused by plant–­parasitic nematodes (PPN) have
been estimated to be $157 billion per year (Abad et al., 2008). Among
these parasites are species of the Meloidogyne genus, also known
as root-­knot nematodes, which are the nematodes that are princi-
pally responsible for damaging horticultural and field crops (Andrés
et al., 2012; Jones et al., 2013). In particular, Meloidogyne incognita
(Kofoid and White) Chitwood has a wide host range, which makes it
one of the most significant plant pathogens (Trudgill & Blok, 2001).

Journal of Phytopathology. 2021;169:229–238. wileyonlinelibrary.com/journal/jph © 2021 Wiley-­VCH GmbH | 229

230 | FERREIRA BARROS et al.
Chemical nematicides are widely used for controlling PPN.
However, due to environmental effects and human toxicity, many
nematicides have been removed from the market, which limits PPN
control options (Burns et al., 2017). Aldicarb[2-­methyl-­2-­(methylthio)
propanal O-­(N-­methylcarbamoyl)oxime] and methyl bromide are ex-
amples of nematicides whose toxicity and environmental effects,
respectively, prompted regulators to ban the chemicals (Norman
et al., 2008; Ruiz-­Suárez et al., 2015). Therefore, there is a great
demand for novel, environmental friendly strategies for controlling
PPN.
Screening plant materials to identify novel compounds is
one suitable approach for finding new nematicidal molecules
(Gommers, 1981). Several volatile compounds from the essen-
tial oils of plants have been shown to possess antimicrobial, in-
secticidal and nematicidal activities (Brahmi et al., 2016; Eloh
et al., 2019; Gupta et al., 2011). Essential oils can be obtained
from several parts of plants and are responsible for their fragrance
(Burt, 2004).
Cinnamomum zeylanicum Blume (syn C. verum), known as true
cinnamon, is a perennial tree that grows in the tropics (Kokate
et al., 1993). It has already been shown to possess fungicidal
and bacterial activities against plant pathogens isolated from ba-
nana, cucumber and watermelon (Choi et al., 2016; Mohammadi
et al., 2016; Ranasinghe et al., 2002). Moreover, C. zeylanicum
essential oil has activity against PPN, including the pine wood
nematode, Bursaphelenchus xylophilus (Kong et al., 2006; Park
et al., 2005). In a recent study, the nematicidal activity of C. zey-
lanicum essential oil against M. incognita was demonstrated by
Eloh et al. (2019) via in vitro assays. However, the authors did not
evaluate the effect of the essential oil and its components in vivo.
Therefore, further studies that aim to evaluate the effect of this oil
and its principal component on greenhouse grown plants should
be carried out. C. zeylanicum essential oil contains several import-
ant compounds including (E)-­cinnamaldehyde, benzaldehyde and
(E)-­cinnamyl acetate with nematicidal potential (Unlu et al., 2010).
However, previous studies assessed effects of compounds that
were different from the chemical assessed here (Kim et al., 2008;
Oka, 2001; Park et al., 2005).
Analogs are molecules that have similar chemical struc-
tures, which can be differentiated by their unique functional
groups. Such compounds may have nematicidal effects; how-
ever, few studies have investigated their potential activities
(Chitwood, 2002; Lopes et al., 2016). Barros, Campos, de Oliveira,
et al. (2019) assessed the nematicidal activity of benzaldehyde
and the oxime analogue of benzaldehyde and found that the
oxime was more efficient than benzaldehyde in vivo. Therefore,
this study aimed to evaluate the toxicity of the essential oil of C.
zeylanicum in juveniles and second-­s tage M. incognita eggs via in
vitro application and greenhouse exposure. Lethal concentrations
(LC 50 and LC95) of the principal component of cinnamon oil, which
was obtained using gas chromatography, and its analog against
M. incognita and the effects of both compounds on egg hatching
were determined.
2 | M ATE R I A L S A N D M E TH O DS
2.1 | Essential oil and nematode inoculum
The essential oil from the leaves of C. zeylanicum, obtained through
steam distillation, was supplied by Brasil Portrait (Sorocaba, São
Paulo, Brazil). Pure M. incognita populations were multiplied on to-
mato plants (Solanum lycopersicum L. 'Santa Clara') and maintained
in a greenhouse for approximately three months. Galled roots
were separated from the soil, washed and cut into segments 0.5-­
cm in length for subsequent egg extractions that were performed
as described by Hussey and Barker (1973). Some extracted eggs
were used for hatching assays, and the remainder were placed in
a hatching chamber and incubated at 28°C. Second-­s tage juve-
niles (J2) that hatched within the first 24 hr of incubation were
discarded, while J2 that hatched throughout the 24–­4 8-­hr period
following the initiation of incubation were used for in vitro and in
vivo experiments.
2.2 | In vitro effect of Cinnamomum zeylanicum on
Meloidogyne incognita J2
Initially, a stock emulsion of C. zeylanicum essential oil in an aque-
ous Tween 80 ® (0.01 g/ml) solution was prepared at a concentra-
tion of 400 µg/ml. This stock emulsion was diluted with 0.01 g/
ml Tween 80 ® to concentrations of 200, 100, 70 and 40 µg/ml.
Then, 500 µl of each final emulsion and 500µl of an aqueous sus-
pension containing 200 M. incognita J2 were added to a 1.5-­m L
microtube, which produced final oil concentrations equal to 100,
50, 35 and 20µg/ml. Tubes containing 0.01 g/ml Tween 80 ® were
used as negative controls. The microtubes were then incubated
at 25°C for 48 hr. Subsequently, samples were homogenized
and 100 μl aliquots were transferred from tubes to the wells of
a 96-­well polypropylene plate, which was used to count mobile
and immobile nematodes under a microscope. Then, one drop of
a freshly prepared 1.0 mol/L NaOH solution was added to each
well, and nematodes were recounted. J2 that changed their body
shape within the 2 min period that followed the addition of NaOH
were considered alive, whereas those whose shapes remained
unchanged were presumed dead (Amaral et al., 2003; Chen &
Dickson, 2000). Data were transformed into percentages prior to
statistical analysis.
2.3 | Infectivity and reproduction of Meloidogyne
incognita J2 exposed to Cinnamomum zeylanicum
essential oil
Initially, a stock solution containing 98 µg/ml C. zeylanicum essen-
tial oil diluted in Tween 80®. Subsequently, in 1.5 ml microtubules, a
500 µl aliquot of the solution was mixed in 500 µl of an aqueous sus-
pension that contained 700 M. incognita J2. The final concentration
FERREIRA BARROS et al.
of essential oil in the microtube was 49 µg/ml, which corresponded
to the concentration needed to kill 50% (LC50) of the nematodes. The
negative control was a 0.01 g/ml Tween 80® solution. As a positive
control, carbofuran (2,3-­dihydro-­2,2-­dimethyl-­1-­benzofuran-­7-­yl-­N
-­methylcarbamate, 98%, Aldrich), a nematicide, was used at a final
concentration (after adding a suspension of J2 in the microtube) of
191 µg/ml, which was determined to be the LC50 of carbofuran in
a study by Terra et al. (2018). The microtubes were stored in an in-
cubator at 25°C for 48 hr. After this period, samples were homog-
enized and 100 μl aliquots were collected and transferred to the
wells of a 96-­well polypropylene plate. J2 mortality was assessed as
previously described. The remaining suspension (900 μl), containing
approximately 630 J2, was added to 4 ml of water and applied to
substrate (Tropstrato®, Vida Verde Indústria e Comércio de Insumos
Orgânicos Ltda) in 300-­ml plastic cups to obtain 30-­d-­old tomato
plants (Solanum lycopersicum L. 'Santa Clara'). The inoculum added
to 4 holes (approximately 2 cm deep) that were mad in the substrate
around the stem. The inoculated plants were kept in a greenhouse
for 35 days. Each root system was removed and washed thoroughly
with still water. After drying with paper towels, root systems were
weighed, galls were counted and eggs were extracted using a tech-
nique described by Hussey and Barker (1973). The eggs were counted
in a Peters chamber under an optical microscope. Numbers of galls
(infectivity) and eggs (reproduction) were calculated per gram of root.
2.4 | In vitro effect of Cinnamomum zeylanicum
essential oil on Meloidogyne incognita J2 hatching
Initially, 1,800, 1,200 and 600 µg/ml stock solutions of C. zeylani-
cum essential oil diluted in Tween 80® were prepared. Subsequently,
500 µl of each essential oil solution and 500 µl of an aqueous sus-
pension that contained 1,000 M. incognita eggs were placed in a
1.5-­ml microtube. Thus, the final oil concentrations of prepared mi-
crotubes were 300, 600 and 900 µg/ml. As negative and positive
controls, a 0.01 g/ml Tween 80® solution and 415 µg/ml carbofuran
solution was used, respectively. After incubation at 25°C for 7 days,
the suspensions were transferred to a counting box and J2 (alive or
dead) that hatched were counted.
2.5 | Effect of Cinnamomum zeylanicum essential
oil solution on Meloidogyne incognita infectivity and
reproduction in tomato plants
Each 25-­day-­old tomato plant (S. lycopersicum L. 'Santa Clara') was in-
dividually transplanted to a 300-­ml plastic cup containing Tropstrato®
substrate. Four mL of 400, 1,000 and 1,600 µg/ml C. zeylanicum es-
sential oil solutions in aqueous Tween 80® (0.01 g/ml) were added
to 4 ml aqueous suspensions containing 700 J2. The final concentra-
tions of oil in resulting suspensions were 200, 500 and 800 µg/ml.
The negative and positive control was comprised of either an aqueous
Tween 80 (0.01 g/ml) solution or 415 µg/ml carbofuran, respectively.
| 231
Inoculations were performed immediately after mixing J2 suspensions
with the essential oil or control solutions. The inoculum was divided
among 4 holes (approximately 2 cm deep) that were dug in substrate
around the stem. Following inoculation, plants were kept in a green-
house for 40 days. Thereafter, each root system was removed from
the substrate, washed with distilled water, dried with a paper towel and
weighed. Then, galls were counted and the number of galls per gram of
root tissue for each sample was calculated.
2.6 | Characterization of the main components of
Cinnamomum zeylanicum essential oil
An analysis of the C. zeylanicum essential oil was performed using
a gas chromatograph–­mass spectrometer (GC-­MS), model QP2010
(Shimadzu Corp.). A 30 m (length) × 0.25 mm (internal diame-
ter) × 0.25 μm (phase thickness) RTX®-­5MS-­Restek capillary column
was employed, and helium was used as a carrier gas with a flow rate
of 1.0 ml/min. The following conditions were adopted: (a) the split/
splitless injector temperature was 220°C; (b) the split ratio was 1:20;
(c) the initial temperature of the column was 60°C; (d) the elevation
rate of the column temperature was 2°C/min up to 200°C and 5°C/
min thereafter; (e) the final temperature of the column was 250°C;
(f) the temperature of the interface between the gas chromatograph
and the mass spectrometer was 220°C; (g) the ionization of each
molecule in the spectrometer resulted from an electron impact of
70 eV; (h) the range of mass/charge (m/z) was 45–­4 00; and (i) the
mass spectrum acquisition time was 0.5 s.
The essential oil was dissolved in acetone to a concentration of
10 mg/ml, and 1 µl was injected into the GC-­MS. A solution of homolo-
gous linear alkanes, containing C9–­C20 carbon atoms, was used as an
external standard. All mass spectra were compared with those in the
NIST 05 Mass Spectral Library (2005), and all peaks in the chromato-
gram with a similarity index <85% were considered unidentified. For
each of the remaining peaks, an arithmetic index (AI) was calculated
according to the following formula: AI = {100Pz + 100[(RT − RTPz)/
(RTPz+1 − RTPz)]}, where Pz was the number of carbon atoms in the
linear alkane with a retention time immediately below that of the sub-
stance to be identified in the chromatogram; RT was the retention time
(min) of the substance to be identified in the chromatogram; RTPz was
the retention time (min) of the linear alkane with a number of carbon
atoms equal to Pz; and RTPz+1 was the retention time (min) of the linear
alkane with a number of carbon atoms equal to Pz+1. Substances with
calculated AI values corresponding to an error ≥3% in relation to those
described by Adams (2007) were considered unidentified.
2.7 | In vitro effect of (E)-­cinnamaldehyde and
(E)-­cinnamaldehyde oxime on Meloidogyne incognita J2
Ninety-­nine per cent pure (E)-­cinnamaldehyde, purchased from Sigma-­
Aldrich, was dissolved in Tween 80® (0.01 g/ml) to a concentration of
600 µg/ml. The resulting solution was diluted with an aqueous solution
232 | FERREIRA BARROS et al.
F I G U R E 1 In vitro effect of C. zeylanicum on M. incognita J2. (a) Experiment 1 (mean of 5 repetitions); (b) Experiment 2, (mean of 5
repetitions). Bars represent the standard error of the mean
of 0.01 g/ml of Tween 80® to concentrations of 500, 400, 300, 200 and
100 µg/ml. Part (500 µl) of each diluted solution was mixed with 500 µl
of an aqueous suspension containing 200 M. incognita J2 in a 1.5-­ml mi-
crotube. The final concentrations of (E)-­cinnamaldehyde produced were
300, 250, 200, 150, 100 and 50 µg/ml. Similarly, (E)-­cinnamaldehyde
oxime (97%) purchased from Sigma-­Aldrich was dissolved in an aqueous
®
solution containing 0.01 g/ml Tween 80 to a concentration of 900 µg/
ml. After samples were diluted and mixed with J2 suspensions, final
concentrations of the oxime were 450, 400, 350, 300, 250 and 200 µg/
ml. An aqueous 0.01 g/ml Tween 80® solution corresponded to concen-
tration equal to zero for both substances. Microtubes were then incu-
bated at 25°C for 48 hr, after which they were homogenized and 100
μL aliquots were transferred to wells of a 96-­well polypropylene plate.
J2 were counted as described previously, and numbers of immobile and
dead J2 were transformed into percentages.
2.8 | Effects of (E)-cinnamaldehyde and
(E)-cinnamaldehyde oxime on Meloidogyne
incognita J2 hatching
An initial 830 and 1,660 µg/ml (E)-­cinnamaldehyde oxime stock so-
®
lutions were prepared in Tween 80 . Subsequently, 500 µl of each
essential oil solution was added to 500 µl of an aqueous suspen-
sion containing 1,000 M. incognita eggs in a 1.5-­ml microtube. Final
(E)-­cinnamaldehyde oxime concentrations were 415 and 830 µg/
®
ml. The negative control was the 0.01 g/ml Tween 80 solution that
was used for diluting (E)-­cinnamaldehyde oxime, while carbofuran
(415 µg/ml) and (E)-­cinnamaldehyde (415 µg/ml) were used as a posi-
tive controls. Following a 7-­day incubation at 25°C, samples were
transferred to a counting box to record the number of J2 that had
hatched (living or dead).
2.9 | Statistics and data analysis
All tests were performed twice, using a completely randomized de-
sign with five replicates per treatment, except for the experiment
that assessed effects of exposing J2 to the LC 50 concentration of
C. zeylanicum essential oil, for which 7 replicates were performed
per treatment. The combined analysis was used for experiments
that showed no difference between the two experiments (p > .05).
Otherwise, we performed a separate analysis for each experimen-
tal repetition. Results were submitted to a normality assessment
(Shapiro–­W ilk test) and the homogeneity of error variance (Bartlett
test) was assessed. When the F test was significant, means were
compared using the Tukey test (p < .05). This test was applied to as-
sess data collected throughout experiments included in ‘Infectivity
and reproduction of M. incognita J2 exposed to the essential oil
of C. zeylanicum’ and ‘In vitro effect of C. zeylanicum essential oil
on M. incognita J2 hatching’ sections of this work. However, to as-
sess experimental data generated in ‘Effect of the essential oil so-
lution of C. zeylanicum on the infectivity and reproduction of M.
incognita in tomato’ and ‘Effects of (E)-­cinnamaldehyde and (E)-­
cinnamaldehyde oxime on M. incognita J2 hatching’ sections, aver-
ages were compared using the Scott and Knott (1974) test (p < .05).
Regarding ‘In vitro effect of C. zeylanicum on M. incognita J2’ and
‘In vitro effect of (E)-­
cinnamaldehyde and (E)-­
cinnamaldehyde
oxime on M. incognita J2’ sections, obtained mortality data were
transformed to percentages and submitted to regression analysis.
To determine the lethal concentration needed to kill 50% and 95%
of the M. incognita J2 assessed (LC 50 and LC95, respectively), per
cent mortality data were submitted to probit analysis. SigmaPlot ®
version 12.0 (Systat Software Inc.) and Sisvar ® (Ferreira, 2019)
software were used to prepare the graphs and perform statistical
analysis, respectively.
FERREIRA BARROS et al. | 233

3 | R E S U LT S negative and positive controls. Compared with the negative control,

hatching was reduced 38%–­54%. Treatment with carbofuran (posi-
3.1 | In vitro effect of Cinnamomum zeylanicum on tive control) did not reduce levels of hatching (Figure 3).
Meloidogyne incognita J2

M. incognita J2 mortality increased with essential oil concentration.
The concentration of essential oil needed to kill 50% of the J2 as-
sessed (LC50) was 56 µg/ml and 41 µg/ml for experiments 1 and 2,
respectively. The concentration of essential oil needed to kill 95% of
the J2 assessed (LC95) was 180 µg/ml and 82 µg/ml for experiments
1 and 2, respectively (Figure 1).
3.2 | Infectivity and reproduction of Meloidogyne
incognita J2 exposed to Cinnamomum zeylanicum
essential oil
When M. incognita J2 were exposed to the LC50 (49 µg/ml) of C. zey-
lanicum essential oil for 48 hr, significant reductions in gall formation
and egg production were observed versus both negative and posi-
tive controls. When compared to the negative control (Tween 80),
exposure to the LC50 (49 µg/ml) of C. zeylanicum essential oil reduced
gall formation and egg production greater than 98%. Compared with
the positive control (carbofuran), exposure to the LC50 (49 µg/ml) of
C. zeylanicum essential oil reduced gall formation and egg production
greater than 97% (Figure 2).
3.4 | Effect of a Cinnamomum zeylanicum essential
oil solution on Meloidogyne incognita infectivity and
reproduction in tomato plants
As a result of treatment with 800 µg/ml C. zeylanicum essential oil,
gall formation was significantly reduced (28%) when compared to
levels produced in the negative control. Carbofuran reduced the
number of galls approximately 87% relative to the number produced
by the negative control (Figure 4).
3.5 | Characterization of the main components of
Cinnamomum zeylanicum essential oil
The main components of C. zeylanicum essential oil were determined
to be (E)-­
cinnamaldehyde (95.1%) and Eugenol (1.8%). The com-
pounds comprised 96.9% of C. zeylanicum essential oil (Table 1).
3.6 | In vitro effect of (E)-­cinnamaldehyde and
(E)-­cinnamaldehyde oxime on Meloidogyne incognita J2

M. incognita J2 mortality increased as the nematodes were ex-

3.3 | In vitro effect of Cinnamomum zeylanicum
essential oil on Meloidogyne incognita J2 hatching
Hatching was significantly reduced by treatment with C. zeylani-
cum essential oil at all concentrations assessed, when compared to
posed to increasing concentrations of E)-­
cinnamaldehyde and
(E)-­cinnamaldehyde oxime (Figure 5). (E)-­cinnamaldehyde oxime
presented was determined to have an LC50 of 323 µg/ml and an LC95
of 529µg/ml, while (E)-­cinnamaldehyde had an LC50 of 64 µg/ml and
an LC95 of 768 µg/ml.

F I G U R E 2 Number of galls and eggs of M. incognita in tomato roots after inoculation of second-­stage juveniles exposed for 48 hr to CL50
(49 µg/ml) obtained for C. zeylanicum. Positive control: Carbofuran 191 µg/ml. Negative control: Tween 80®. (a) Experiment 1 (mean of 7
repetitions); (b) Experiment 2 (mean of 7 repetitions). Means with the same lowercase letter (galls) and uppercase letter (eggs) do not differ
significantly (p < .05) by the Tukey test. Bars represent the mean standard error [Colour figure can be viewed at wileyonlinelibrary.com]
234 | FERREIRA BARROS et al.
F I G U R E 3 Hatching of M. incognita J2 from eggs exposed to
300, 600 and 900 µg/ml. C. zeylanicum essential oil. As positive
and negative controls, 415 µg/ml carbofuran and Tween 80® were
used, respectively. Means with the same lowercase and uppercase
letters do not differ significantly (p < .05) via the Tukey test. Bars
represent the standard error of the mean [Colour figure can be
viewed at wileyonlinelibrary.com]
F I G U R E 4 The number of galls produced after inoculation of M.
incognita J2 with different concentrations of C. zeylanicum essential
oil. As positive and negative controls, 415 µg/ml carbofuran
and Tween 80® were used, respectively. Means with the same
lowercase and uppercase letters do not differ significantly (p < .05)
via the Tukey test. Bars represent the standard error of the mean
[Colour figure can be viewed at wileyonlinelibrary.com]
TA B L E 1 Principal components of C. zeylanicum essential oil
determined via gas chromatograph–­mass spectrometry (GC-­MS)
Área
Component RT (min) AI Si (%) (%)
(E)-­Cinnamaldehyde 22.495 1.273 97 95.1
Eugenol 27.644 1.358 94 1.8
Total -­-­ -­-­ -­-­ 96.9
Abbreviations: RT, retention time; AI, calculated arithmetic index; SI,
similarity index.
3.7 | Effects of (E)-­cinnamaldehyde and
(E)-­cinnamaldehyde oxime on Meloidogyne incognita
J2 hatching
Both (E)-­
cinnamaldehyde and its oxime significantly reduced J2
hatching when compared to negative and positive controls. The
positive control (carbofuran) did not differ from the negative con-
trol. At a concentration of 415 µg/ml, (E)-­cinnamaldehyde and (E)-­
cinnamaldehyde oxime similarly reduced levels of J2 hatching. The
greatest reduction in hatching (77%) was observed in nematodes
exposed to 830 µg/ml (E)-­cinnamaldehyde oxime (Figure 6).
4 | D I S CU S S I O N
Plant essential oils and their constituents have the potential to
be used to control nematode populations and can be used as ne-
maticides. In addition to compounds derived from oils, their re-
spective analogs may also possess nematicidal activity, which may
have enhanced activity relative to oil derived compounds (Andrés
et al., 2012; Barros, Campos, Paula, et al., 2019). The C. zeylanicum
essential oil showed strong toxicity in M. incognita activity. After
48 hr of direct contact with J2, the average LC50/48 hr value for C.
zeylanicum essential oil was 49 µg/ml. Eloh et al. (2019) determined
that the LC50/24 hr of C. zeylanicum essential oil against M. incog-
nita was 391 ± 92 µg/ml. Kong et al. (2006) determined that the
LC50/24 hr of cinnamon bark oil against Bursaphelenchus xylophilus
was 120 µg/ml. Further, LC50/72 hr values for cinnamon leaf oil and
cinnamon bark oil against M. graminicola in rice root galls were 0.326
and 0.454 µg/ml, respectively (Amarasinghe et al., 2011). Although
the nematicidal activity of C. zeylanicum has been studied, the activ-
ity measured in galls, eggs and hatching has not been demonstrated.
In this study, when J2 were exposed to the LC50 of C. zeylani-
cum essential oil and subsequently used to inoculate tomato plants,
quantities of galls and eggs were greatly reduced. This reduction
was superior to that of the nematicide carbofuran, which was a
well-­
known nematicide that was widely used before its restric-
tion. Barros, Campos, Paula, et al. (2019) also observed that num-
bers of galls and eggs were greatly reduced when M. incognita J2
were exposed to Dysphania ambrosioides essential oil. Further, the
FERREIRA BARROS et al.
F I G U R E 5 In vitro effect of (E)-­cinnamaldehyde and (E)-­cinnamaldehyde oxime treatment on M. incognita J2. (a) Effects of (E)-­
cinnamaldehyde oxime are shown (Mean of 10 repetitions). (b) Effects of (E)-­cinnamaldehyde (Mean of 10 repetitions) are shown. Bars
represent the standard error of the mean
F I G U R E 6 M. incognita J2 hatching from eggs treated with
(E)-­cinnamaldehyde and (E)-­cinnamaldehyde oxime. As positive
and negative controls, 415 µg/ml carbofuran and Tween 80® were
used, respectively. Means with the same lowercase and uppercase
letters do not differ significantly (p < .05) via the Scott–­Knott test.
Bars represent the standard error of the mean [Colour figure can be
viewed at wileyonlinelibrary.com]
researchers determined that 50% of the nematodes that remained
alive postexposure likely suffered from nervous system defects that
had the potential to prevent host recognition, which is a phenome-
non that was also likely observed in this study.
Significant reductions in levels of M. incognita hatching were also
observed after exposure to all concentrations of C. zeylanicum essen-
tial oil assessed. Carbofuran, on the other hand, did not affect levels
| 235
of M. incognita hatching. Similar findings were determined by Jardim
et al. (2018) who assessed the effectiveness of Cinnamomum cassia
oil against M. incognita hatching. The inability of carbofuran to inhibit
M. incognita hatching has been demonstrated previously (Barros,
Campos, Paula, et al., 2019; Pedroso et al., 2019; Silva et al., 2019).
Several factors affect the chemical composition of C. zeylani-
cum essential oil. These include the part of the plant from which it
is extracted, age of trees, season, place of cultivation and extraction
method used (Haddi et al., 2017). The main component of cinnamon
oil is variable. In this work, the main components of cinnamon essen-
tial oil were (E)-­cinnamaldehyde and eugenol, and the same finding
was obtained by Wang et al. (2009) in oil extracted from cinnamon
leaves. According to López-­Serna et al. (2016), cinnamaldehyde is a
highly volatile substance and is easily oxidized to cinnamic acid upon
exposure to air. Therefore, soil composition, including its microbi-
ota, may influence cinnamaldehyde volatility and degradation, which
may explain why only a minor reduction in the number of galls was
observed when essential oil emulsions were placed on J2-­infested
substrates in this work.
The efficacy of (E)-­
cinnamaldehyde and (E)-­cinnamaldehyde
oxime was evaluated in the present study. The LC50/48 hr of
(E)-­
cinnamaldehyde was determined to be 64 µg/ml, while the
LC50/48 hr of (E)-­cinnamaldehyde oxime was 323 µg/ml. Although
(E)-­
cinnamaldehyde oxime had a greater LC50 value than (E)-­
cinnamaldehyde, oximes are generally advantageous since they are
typically highly stable and can be easily prepared from several func-
tional groups including aldehydes and ketones (Corsaro et al., 2009).
Cinnamaldehyde oxime represents a new nitric oxide donor and it
has the potential to be a very important tool for future prevention
or treatment of conditions where nitric oxide production is deficient,
such as atherosclerosis, hypertension, cardiac ischaemia and throm-
bus formation, which indicates its low toxic effect to humans (Veras
et al., 2013).
236 | FERREIRA BARROS et al.
The activities of oximes of other substances have previously
been evaluated against M. incognita. For example, benzaldehyde
oxime more greatly reduced galls and eggs in tomato roots than
benzaldehyde (Barros, Campos, de Oliveira, et al., 2019). Oximes
have low volatility and, consequently, greater stability. This allows
the molecules to remain in the soil for a long period of time, which
enhances their ability to prevent nematode growth. Another study
determined that an analogue of 9H purine and two analogs of di-
hydrouracil could control M. incognita as efficiently as the nemati-
cide carbofuran (Lopes et al., 2016). Thus, it is potentially useful to
obtain analogues of cinnamaldehyde, which may possess similar or
increased nematicidal activities and greater stability than the cin-
namaldehyde molecule itself.
Both substances [(E)-­cinnamaldehyde and (E)-­cinnamaldehyde
oxime] similarly reduced levels of hatching. One experiment re-
vealed that, at a concentration of 830 µg/ml, cinnamaldehyde oxime
most efficiently reduced levels of hatching compared with the other
compounds and concentrations assessed. Other substances isolated
from essential plant oils, such as 2-­undecanone, ascaridol, carvac-
rol, thymol and linalool, have already been reported to have ovicidal
effects (Faria et al., 2016; Ibrahim et al., 2006). As mentioned pre-
viously, the failure of carbofuran to reduce levels of hatching is in
accordance with findings of several studies (Barros, Campos, Paula,
et al., 2019; Pedroso et al., 2019; Silva et al., 2019). The lipid layer
present in the nematode eggshell is the principal barrier of chemical
permeability (Gaugler & Bilgrami, 2004). Due to this layer, many sub-
stances are unable to interfere with egg hatching. Therefore, the re-
sults obtained support the idea that cinnamaldehyde oxime is a good
option for M. incognita management. However, more studies aimed
to determine the precise mechanism of cinnamaldehyde oxime func-
tioning are required.
AC K N OW L E D G M E N T S
The authors gratefully acknowledge financial support and fellow
ships from: Fundação de Amparo à Pesquisa do Estado de Minas
Gerais (FAPEMIG), Coordenação de Aperfeiçoamento de Pessoal de
Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento
Científico e Tecnológico (CNPq).
C O N FL I C T O F I N T E R E S T
The authors declare no conflict of interest.
AU T H O R C O N T R I B U T I O N S
AFB, VPC, DFO and JCPS conceived and designed the study. AFB,
LLP, LAP and FJS conducted bioassays. GHS carried out the GC–­MS
analysis and interpreted the results. All authors contributed to in-
terpreting the results, reviewed the manuscript and approved the
manuscript.
PEER REVIEW
The peer review history for this article is available at https://publo​
ns.com/publo​n/10.1111/jph.12979.
DATA AVA I L A B I L I T Y S TAT E M E N T
The data that support the figures of this study are available from the
corresponding author upon reasonable request.
ORCID
Aline Ferreira Barros https://orcid.org/0000-0003-0461-802X
Vicente Paulo Campos https://orcid.org/0000-0002-3284-7412
Letícia Lopes de Paula https://orcid.org/0000-0002-9735-0036
Luma Alaís Pedroso https://orcid.org/0000-0001-9964-1585
Fabíola de Jesus Silva https://orcid.org/0000-0002-1734-6521
Júlio Carlos Pereira da Silva https://orcid.
org/0000-0002-1961-6695
Denilson Ferreira de Oliveira https://orcid.
org/0000-0001-9326-8716
Geraldo Humberto Silva https://orcid.
org/0000-0001-7928-8980
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How to cite this article: Ferreira Barros A, Paulo Campos V,
Lopes de Paula L, et al. The role of Cinnamomum zeylanicum
essential oil, (E)-­cinnamaldehyde and (E)-­cinnamaldehyde
oxime in the control of meloidogyne incognita. J Phytopathol.
2021;169:229–­238. https://doi.org/10.1111/jph.12979