Keywords:

Antimicrobial activity

Biocontrol

Plant extracts

Medicinal plant

Sodium chloride

Effect of salinity levels on antifungal activity of essential oil from Thymus against Fusarium

oxysporum

Efecto de los niveles de salinidad sobre la actividad antifúngica del aceite esencial de Thymus contra

Fusarium oxysporum

Efeito dos níveis de salinidade na atividade antifúngica do óleo essencial de Thymus contra Fusarium

oxysporum

Boualem Boumaaza

1,2

M’hamed Benada

Soane Boudalia

Ibrahim E. Benzohra

Abdelhamid Gacemi

Omar Khaladi

Mohamed Benkhelifa

Rev. Fac. Agron. (LUZ). 2022, 39(3): e223941

ISSN 2477-9407

DOI: https://doi.org/10.47280/RevFacAgron(LUZ).v39.n3.07

Crop Production

Associate editor: Dra. Lilia Urdaneta

University of Zulia, Faculty of Agronomy

Bolivarian Republic of Venezuela

Department of Agronomy, University of Abdelhamid Ibn

Badis, BP 300, 27000, Mostaganem, Algeria.

University of Ibn khaldounTiaret. BP 78 zaâroura 14000,

Tiaret, Algéria.

University of Ahmed Zabana. BP 48000 Bormadia Relizane.

Laboratory of Biology, Water and Environment (LBEE),

University of 8th May, 1945 Guelma, BP 401 24000 Guelma,

Algeria.

Scientic and Technical Research Center on Arid Regions

(CRSTRA) BP. Box 1682 RP 07000, Biskra, Algeria.

Conservation Laboratory Wetlands.

INRA Station de Recherche Hmadna. BP 48017 Relizane.

Received: 01-06-2022

Accepted: 02-08-2022

Published: 22-082022

Abstract

Thyme (Thymus sp.), a medicinal plant of the family Lamiaceae, is

used in traditional medicine, contains a wide array of medicinally active

components, in their great majority of a rather complex mixture of thymol,

ρ-Cymene, γ-Terpinene, β-Caryophyllen, etc. This study aimed to evaluate

the efcacy of Thymus vulgaris extract against Fusarium oxysporum f. sp.

radicis-lycopersici strain under saline conditions, assuming soil with high salt

content of the arid regions. Essential oil was extracted by hydrodistillation

technique using a Clevenger apparatus. The essential oil compounds were

identied by GC-MS analysis. Antifungal activity of essential oil against

Fusarium oxysporum f. sp. radicis-lycopersici was investigated by agar

dilution method. The main constituents of thyme essential oil were thymol

(76.96 %), ρ-cymene (9.89 %) and γ-Terpinene (1.92 %). Essential oil from

Thymus resented high in vitro activity, in controlling conidial germination

and mycelial growth. However, the oil was signicantly not active against

the spore production under a salinity medium. The results showed that

mycelial growth was stimulated in concentrations with 0.6-1.5 %. In

contrast, it was signicantly reduced at a higher concentration (2 %). The

application of NaCl caused a signicant increase in the conidia production

at various concentrations tested. NaCl has a minor inhibitory effect on

conidial germination only when the concentration was 2 %. The results of

this study indicate that salinity decreases the efcacy of essential oil against

the pathogen.

This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.

Rev. Fac. Agron. (LUZ). 2022, 39(3): e223941. July - September. ISSN 2477-9407.

2-6 |

Resumen

El tomillo (Thymus sp.), una planta de la familia Lamiaceae,

se utiliza en la medicina tradicional, contiene una amplia gama

de componentes medicinalmente activos, en su gran mayoría de

una mezcla bastante compleja de timol, ρ-cimeno, γ-terpineno y

β-Carioleno, entre otros. El objetivo del presente estudio fue evaluar

la ecacia del extracto de Thymus vulgaris contra Fusarium oxysporum

f. sp. radicis-lycopersici bajo condiciones salinas, simulando suelos

con altos contenidos de sales de las regiones áridas. El aceite esencial

se extrajo mediante hidrodestilación utilizando el aparato Clevenger.

Los compuestos del aceite esencial se identicaron mediante el análisis

GC-MS. La actividad antifúngica del aceite se evaluó mediante el

método de dilución en agar. Los principales componentes del aceite

esencial del tomillo fueron timol (76,96 %), ρ-cimeno (9,89 %) y

γ-terpineno (1,92 %). El aceite presentó alta actividad in vitro, en el

control de la germinación de los conidios y del crecimiento micelial.

Sin embargo, el aceite no inhibió la esporulación en el medio salino.

El crecimiento micelial fue estimulado al 0,6-1,5 %, pero se redujo

signicativamente a la concentración más alta (2 %). La aplicación de

NaCl provocó un aumento signicativo en la producción de conidios

en todas las concentraciones. La germinación conidial fue inhibida

ligeramente por el NaCl solo al 2 %. Los resultados indican que la

salinidad disminuye el potencial antifúngico del aceite esencial de

Thymus contra el patógeno.

Palabras clave: Actividad antimicrobiana, biocontrol, extractos de

plantas, planta medicinal, cloruro de sodio.

Resumo

O tomilho (Thymus sp.), planta medicinal da família Lamiaceae,

utilizada na medicina tradicional, contém uma grande variedade de

componentes medicinais ativos, em sua grande maioria uma mistura

bastante complexa de Timol, ρ-Cymene, γ-Terpinene, β-Caryophyllen,

etc. O presente estudo foi avaliar a ecácia do extrato de Thymus

vulgaris contra Fusarium oxysporum f. sp. radicis-lycopersici sob

condições salinas, assumindo solos com alto teor de sal das regiões

áridas. O óleo essencial por extraído pe la técnica de hidrodestilação

utilizando aparelho de Clevenger. O composto de óleo essencial por

identicado por análise GC-MS. Atividade antifúngica do óleo esencial

contra Fusarium oxysporum f. sp. radicis-lycopersici por investigado

pelo método de diluição em agar. O principais constituintes do óleo

esencial de tomilho foram timol (76.96 %), ρ-cimeno (9.89 %) e

γ-Terpineno (1.92 %). O óleo esencial de Timo apresentou alta ativida

de in vitro, no control da germinação de conídios e do crescimento

micelial. No entanto, o óleo não por signicativamente ativo contra

produção de esporos meio de salinidade. Os resultados sugerem que

o crescimento micelial por estimulado em concentrações de 0,6-1.5

%. Pelo contrário, reduziu signicativamente na concentração mais

alta (2 %). A aplicação de NaCl causou um aumento signicativo

na produção de conidiogênese em várias concentrações testadas. A

germinação dos conídios for levemente inibida pelo cloreto de sódio

somente quando a concentração for de 2 %. Os resultados este estudo

indicaque a salinidade diminui a ecácia do óleo esencialcontra o

patógeno.

Palavras-chave: Actividad antimicrobiana, biocontrole, extratos de

plantas, planta medicinal, cloreto de sódio.

Introduction

Fusarium oxysporum is an ubiquitous fungus, widely distributed

among soil, plants, plant debris and other organic substrates. This

fungus is one of the most important phytopathogenic and toxigenic

fungi, affecting the health and survivability of plants in more than

100 different crops (Al-Hatmi et al., 2016). Additionally, Fusarium

oxysporum has numerous specialized forms (f. sp.) that infect a

range of host plants causing diseases such as vascular wilt, corm

rot, and root rot (Edel-Hermann and Lecomte, 2019). Another

important characteristic of the species of this genus is their ability to

produce a various of mycotoxins that can play an important role in

the pathogenesis (Lombard et al., 2019). Preventive measures, such

as chemical pesticides have controlled the disease to some extent.

However, excessive application of fungicides has led to severe damage

to soil microbial communities and fertility and may leave behind

toxic residues in treated products (Meena et al., 2020). Additionally,

intensive use of fungicides can result in the development of pathogens

acquired resistance to the fungicide (Corkley et al., 2021).

Due to the problems caused by chemical substances, the

development of alternative control measures is of great importance.

Biological control has been considered as a desirable and realistic

alternative. Numerous studies have demonstrated the ability of

several essential oils to possess antibacterial, antifungal, antiviral,

antioxidant activities and play an important role in the protection of

the plants against plant pathogens both in vitro and in vivo (Mutlu-

Ingok et al., 2020; Marín-Tinoco et al., 2021). Note that essential oils

can be an effective solution, as their toxicity is much lower, better

specicity of action, biodegradable and environmentally friendly

(Campos et al., 2019). Being volatile, they can act as fumigants,

repellents, and contact insecticides or as reproduction inhibitors (Gao

et al., 2020; Silva-Marrufo and Marín-Tinoco, 2021). Essential oils

interact with the odor receptors of insect pests (Ceratitis capitata),

triggering various behaviors: ight, attraction, oviposition, etc.

(Benelli et al., 2012). The studies published on the activity of

essential oils as herbicides are numerous and generally cover seed

germination inhibition tests (Yilar et al., 2020). Those that appear

to be the most active are essential oils containing phenols (thymol,

carvacrol), ketones (carvone, pulegone) or etheroxides (eucalyptol or

1,8-cineol).

Plants of the genus Thymus

are widely used in food avouring,

as well as aroma additives, in perfumery, in folk medicine and in

pharmacological sector as natural antioxidants and antimicrobial

agents (Gema, 2020). In North Africa, most of the thyme produced

is in the form of dried herbs. With mean annual rainfalls of 150-400

mm.years

-1

, the yield of Thymus vulgaris as fresh herbs may be 5 to 6

t.ha

-1

with 2 t.ha

-1

for the production of dried herbs. With an irrigated

system, in the presence of fertilization, the yield can be 9.77 t.ha

-1

with an oil recovery yield of 43.14 to 48.9 kg.ha

-1

(Kozera et al.,

2015).

It has been reported that essential oils possess various actions on

microorganisms, such as disruption of the cytoplasmic membrane,

disruption of the proton driving force, electron leakage and

coagulation of the protein content of cells, acidication from inside

the cell, blocking the production of cellular energy and the synthesis

of structural components. Thymol and carvacrol have antimicrobial

activity, which is related to inhibition of membrane permeability,

resulting in ions and ATP leakage, inhibition ergosterol biosynthesis

and cell death (Alizadeh et al., 2018).

This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.

3-6 |

Antimicrobial activity of essential oils is strongly affected by

many factors such as high temperature, poor water solubility, light,

and oxygen (de Souza et al., 2008; Turek and Stintzing, 2012). This

study aimed to assess the antifungal activity of the essential oil from

Thymus against Fusarium oxysporum f. sp. radicis-lycopersici in

growth media amended with various levels of sodium chloride.

Materials and methods

Inoculum preparation

The tested Fusarium oxysporum f. sp. radicis-lycopersici strain

M27b27245 was preserved at the Plant Protection Laboratory of the

Agronomy Science Department, University of Abdelhamid Ibn Badis,

Mostaganem, Algeria. Fusarium oxysporum readily produced conidia

after 14 days on potato dextrose agar (PDA; potato dextrose agar.

SPA CRAPC Algeria) plate at 25

C in the dark. Conidial suspension

was obtained by ooding plates with distilled water and rubbing

gently with a glass rod, then ltering through sterile cheesecloth.

The conidia concentration present in the initial suspension (adjusted

to the concentration 1x10

conidia.mL

-1

) was quantied using the

hemocytometer.

Analysis of the essential oil of Thymus vulgaris

Thymus vulgaris was supplied for the study by the National

Agronomic Institute of Tunisia (INAT), and the experiment

was conducted from December 2017 to January 2018. Hydro-

distillationwascarried out using a Clevenger-type device according

to the method recommended in British Pharmacopoeia (British

Pharmacopoeia, 2016), where 500 g of vegetable matter is introduced

with 3 L water into a 5 L ask. After installation and closing of the

assembly, the ask heater is started up with an optimum adjustment of

the heating to allow stability of the extraction at a constant and well-

controlled speed. The vapor charged with essential oil arrives in the

condenser. The total duration of the extraction was estimated at 3 h.

The essential oil differs from the hydrosol by its difference in density

and color. It is separated from the by decantation. It is then dried with

anhydrous sodium sulfate (Na

) then recovered and stored in a

cool place (4 °C).

Oil extracted was analyzed by GC/MS using a gas chromatograph

(HP 5890-Serie II) coupled to a mass spectrometer (HP-MSD 5972 A)

with an HP-5MS capillary column (30 m x 0. 25 mm, lm thickness,

0.25 µm). The temperature of the column was programmed at 50

for 1 min, then 7

C / min at 250

C, and nally left at 250

C for 5 min.

The helium injection (1 mL.min

-1

). The temperature of the injector

port was 240

C, while that of the detector set at 250

C. The ionizing

energy was 70 eV, the full-scan mass spectra: from 40 to 500 amu.

The identication of the components was carried out on the basis of

chromatographic retention indices and by comparison of the mass

spectra recorded with the calculated spectral library (Al-Asmari et

al., 2017).

In vitro antifungal assay

Effects on mycelial growth

The antifungal activity of the essential oil against to Fusarium

oxysporum f. sp. radicis-lycopersici was investigated by agar dilution

method (Souza et al., 2002). In sterile Petri dishes with 9 cm diameter

containing 20 mL potato dextrose agar (PDA) mixed with ve levels

of NaCl to obtain nal concentrations 0.3, 0.6, 1, 1.5 and 2 %.

Then, 100 μL of the essential oil was evenly spread on the surface

of Petri dishes containing PDA + NaCl. Essential oil was previously

diluted, to then adjust the concentration to 1 %. 30 μL of the fungi

Boumaaza et al. Rev. Fac. Agron. (LUZ). 2022, 39(3): e223941

suspension 105 conidia.mL

was placed at the center of each Petri

plate. In this assay, an amount of essential oil was added at minimum

inhibitory concentration (MIC) (1 %). Cultures were incubated at 25

°C in the dark. Colony diameters were measured in two perpendicular

directions, after 24 h and again after 96 h. Each treatment had ve

replications. In the control group, sterile distilled water was used in

place of the oil, and no additional NaCl was added.

Effects on sporulation

Conidia suspension of Fusarium oxysporum f. sp. radicis-

lycopersici was obtained from two weeks-old culture incubated at 25

C in the dark. Plates were ooded with sterile-distilled water using

0.5 % (v/v) Tween 80 solution and conidia were gently dislodged

from the mycelium using a sterile magnetic stir bar placed on the

agar and set stirring for 5 min to loosen the conidia. The suspension

was ltered through a glass lter to eliminate mycelium. Finally, the

conidia concentration was quantied using the hemocytometer.

Effects on the germination of conidia

To determine the inuence of NaCl upon the antifungal activity

of the essential oil on spore germination of Fusarium oxysporum f.

sp. radicis-lycopersici, a drop containing 100 conidia was transferred

on to water agar plates enriched with NaCl and essential oil. The

plates were incubated at 25

C in the dark for 24 h. The results were

expressed as the percentage of germinated conidia observed under

the optical microscope at 400x magnication. The conidium was

considered as germinated if the germ tube length was at least twice

the length of the conidium.

All results are presented as mean ± standard deviation. Results

were analyzed using at test and analysis of variance (ANOVA).

Signicance was considered at P< 0.05 using StatBox®, Version 6.0.4

(Grimmer Soft, Fr.) (2009). The tests were performed in triplicate.

Results and discussion

Essential oil chemical composition

Seventeen compounds of the essential oil of Thymus vulgaris

representing 99.24 % of the total detected constituents were identied

(table 1). Among the mentioned phenolic compounds, thymol was

present in the largest amounts (76.69 %) followed by ρ-cymene (9.89

%), γ-terpinene (1.92 %); caryophyllene oxide (1.69 %); β-linalool

(1.51 %) and β-caryophyllen (1.32 %). A similar result was previously

described by Kolsum et al. (2017) that showed 32 compounds from

Thymus vulgaris with dominance of thymol (32.67 %) and p-cymene

(16.68 %).The major components of the oil were thymol (36.81 %)

and p-cymene (30.9 %) (Moghaddam and Mehdizadeh, 2020).

Activity of essential oil on mycelial growth

The results of antifungal activity assays showed that the 1 % (v/v)

thyme essential oil signicantly inhibited (P<0.01) the mycelium

growth of Fusarium oxysporum f. sp. radicis-lycopersici after 24 h of

incubation at 25

C compared to untreated (gure 1a).

The essential oil reached an inhibition rate of 43 % for Fusarium

oxysporum f. sp. radicis-lycopersici after culturing for 48 h. In the

research carried out by Galovičová et al. (2021), amounts of free

thyme oil between 62.5 and 500 μL.mL

-1

were necessary to inhibit 46-

87 % the growth of Serratia marcescens. Thymus vulgaris essential

oil completely suppressed the colony growth of Fusarium oxysporum

f. sp. radicis-lycopersici (Kumar et al., 2007; Aksit et al., 2022). After

96 h, the mycelium growth of the strain on the essential oil-treated

groups became larger, and the inhibition rate declined (10.86 %),

which may be because the oil had evaporated.

This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.

Rev. Fac. Agron. (LUZ). 2022, 39(3): e223941. July - September. ISSN 2477-9407.

4-6 |

Table 1. Chemical constituents of Thymus vulgaris leaves

essential oil using gas chromatography-mass

spectrometry (GC/MS).

The opposite result was observed for essential oil from Thymus.

In which conidia production of Fusarium oxysporum f. sp. radicis-

lycopersici was stimulated (gure 2). The average number of

conidia on unamended (control) PDA media plates after 14 days

of incubation at 25

C were 5.65x10

conidia.mL

-1

while grown

colonies amended with Thymus essential oil, production of conidia

was signicantly increased than the control, and up to 13.7x10

conidia.mL

-1

. Thymol and p-cymene are the major compounds

found in Thymus vulgaris and have shown potent fungicidal and/

or fungistatic activities against various phytopathogenic fungi such

as Fusarium spp., Aspergillus spp., Cladosporium spp., Mucor spp.

and Rhizopus spp. (Kumar et al., 2007). Thymol and p-cymene

provide irreversible damage to the cell wall, cytoplasm membrane

and nuclear membrane of

lamentous fungi (Rasooliand Owlia,

2005).

The effects of essential oil on the spore germination of Fusarium

oxysporum f. sp. radicis-lycopersiciare shown in gure 3. In the

absence of NaCl, thyme oil was found to be the most effective

inhibition (86 %) of spore germination. The essential oil may

inhibit the cell growth and proliferation by interrupting ergosterol

biosynthesis (Gao et al., 2016). Essential oil enables us to integrate

into the lipids of the cell membrane, increasing permeability occur as

a due of loss of ions and reduction of membrane potential, collapse

of the proton pump and depletion of the ATP pool, which eventually

lead to leaking of intracellular constituents, coagulation of cell

contents, lysis and cell death (Turgis et al., 2012). Antimicrobial

activity was due to disturbance of the permeability of the membrane

and the release of some inhibition enzymes such as ATPase, histidine

decarboxylase, and amylase from the cellular content (Scollard et al.,

2016). Carvacroland thymol disrupted the ergosterol biosynthesis

and membrane integrity (Ahmad et al., 2011). Evaluation of Thymus

vulgaris essential oil against fungi has shown a decrease from 4.04

to 6.27 fold of Tri4 gene expression by the use of qRT-PCRassay

(Kolsum et al., 2017).

Figure 1. Effect of salinity levels and essential oil on mycelial

growth of Fusarium oxysporum at 24, 48, 72 and 96

h. (1a) Effect of fungal load of essential oil. (1b) Effect

of salinity on mycelial growth. (1c) Effect of salinity and

essential oil on mycelial growth. ns, *, ** and *** non-

signicant, signicant at 0.05, 0.01 and 0.001 probability

level, respectively.

Compounds RT.min-

RI Content (%)

Thujene

α-Pinene

cis-Sabinene hydrate

Camphene

β-Myrcene

α-Terpinene

β- Phelladrene

γ-Terpinene

ρ-Cymene

Carvacryl Acetate

β-Linalool

4-Terpineol

β-Caryophyllen

Borneol

Caryophyllene oxide

Thymol

Eugenol

10.24

10.32

10.66

10.76

11.74

11.88

12.18

12.55

12.80

13.72

15.36

16.07

16.12

17.19

20.18

21.99

21.61

1140

1165

777

846

889

916

1012

965

1024

1303

998

1107

1319

1012

1479

1196

1300

0.71

0.49

0.46

0.14

0.71

0.58

0.37

1.92

9.89

1.19

1.51

0.65

1.32

0.49

1.69

76.96

0.16

Total identied (%) 99.24

(a) Effect of fungal load of essential oil

(b) Effect of salinity levels (0.3, 0.6, 1, 1.5 and 2 %)

Figure 2. Effect of salinity and essential oil on conidia production

by Fusarium oxysporum. Values with different letters

show signicant difference (P≤0.05).

This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.

Boumaaza et al. Rev. Fac. Agron. (LUZ). 2022, 39(3): e2239415-6 |

Effects of NaCl on mycelial growth, germination and conidia

production

After 24 h of incubation, the mycelial growth eached a diameter

of 0.82 mm in the control. Whereas under gradual concentrations of

NaCl, colony diameters were 1.2 mm at 0.3 % NaCl, 1.27 mm at 0.6

%, 1.3 mm at 1 %, 1.35 mm at 1.5 % and 1.2 mm at 2 % respectively

(gure 1b). The average mycelial growth was signicantly in the

saline medium over the ve levels of NaCl compared to the control.

Figure 1b shows that the saline medium has a signicant inuence on

the stimulation of mycelial growth compared to the control. Similar

supporting results have been reported (Boumaaza et al., 2015).

Figure 2 shows that the application of sodium salt caused a

signicant increase in the conidia production at various concentrations

tested compared with control (P< 0.001). By adding 2 % of NaCl

to the culture medium, an increase in spore production by 13.6x10

conidia.mL

-1

can be obtained compared to the control 5.65x10

conidia.mL

-1

. Ahigh concentration of sodium chloride showed to be

the most favorable to the sporulation.

The germination of conidia that were affected by higher levels of

NaCl is shown in gure 3. An inhibition of 2.66 % in germination was

observed at 2 % salinity. However, the lower levels of NaCl (0.3, 0.6

and 1 %) were signicantly not active against Fusarium oxysporum f.

sp. radicis-lycopersici.

Figure 3. Effect of salinity and essential oil on conidia germination

by Fusarium oxysporum. Values with different letters

show signicant differences (P≤0.05).

Combined effects of salinity and essential oil on Fusarium

oxysporum

The combined effects of salinity levels and essential oil on

Fusarium oxysporum are shown in gure 1c. After 48h of incubation,

the growth of the mycelium treated with essential oil decreased to

higher salinity levels. Compared to untreated salinity (3.4 mm), the

mycelium growth achieved 3.63, 3.68, 3.26, 3.41 and 2.48 mm was

obtained by the concentrations of 0.3, 0.6, 1, 1.5 and 2 % NaCl,

respectively.

The results shown in gure 2 revealed that the combined effects of

salinity levels and essential oil caused a signicant increase in spore

production at various levels tested compared with control. However,

this combination increase the antifungal activity of essential oil

compared to oil alone (13.7x10

conidia.mL

-1

), the conidia production

achieved 2.66, 3.5, 3.6, 5.2 and 9.76x10

conidia.mL

-1

were obtained

by with concentrations of NaCl 0.3, 0.6, 1, 1.5 and 2 %, respectively.

As seen from gure 3, combined effects of salinity levels and

essential oil caused a signicant decrease in the antifungal activity

of essential oil compared to oil alone (14 %). The spore germination

achieved of 20.66, 57, 58.66 and 58 % was obtained by the

concentrations of 0.3, 0.6, 1 and 1.5 % respectively. The results of

this study indicated that salinity decreases the efcacy of essential

oils against the pathogen.

According to Perumal et al. (2016), the efcacy of essential oils

against the microorganisms mainly depend on the characteristics

of their components and inuenced by many factors such as high

temperature, low water activity, low nutrient conditions and UV light

for effective establishment and disease control. Several studies have

investigated the control efcacy of essential oils against Fusarium sp.

(Xing et al., 2014). To date, there have been limited studies on the use

essential oils in mycology under salinity conditions. In accordance

with Biswas et al. (2011), essential oil contents are also affected by

environmental factors such as salinity, water stress and soil pollution

(Stancheva et al., 2014). The degradation of essential oils depends

on some climatic and edaphic factors that inuence the course of

antifungal activity. The constituents of essential oils are particularly

prone to oxidative damage; like light and heat, salts, especially copper

and ferrous ions, are thought to promote auto-oxidation (Choe and

Min, 2006).

Conclusion

According to these results, the essential oil of thyme showed

signicant antifungal activity on the strain studied. Also, it can be said

that the salinity of the culture medium leads to an in vitro stimulation

of mycelial growth, there by reducing the antifungal activity of this

oil against this pathogen. In the absence of NaCl, thyme oil was found

to be the most effective inhibition of spore germination.

Literature cited

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