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Antifungal activity of Moroccan plants against Penicillium italicum, the causal

agent of blue mold of citrus

Conference Paper · March 2010

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 Integrated Control in Citrus Fruit Crops
IOBC/wprs Bulletin Vol. 62, 2011
pp. 51-54

Antifungal activity of Moroccan plants against Penicillium italicum,

the causal agent of blue mold of citrus

L. Askarne, I. Talibi, M. A. Serghini, H. Boubaker, F. Msanda, A. Ait Ben Aoumar,

E. H. Boudyach
Laboratoire de Biotechnologies et Valorisation des Ressources Naturelles, équipe de
phytopathologie et lutte intégrée, Université Ibn Zohr, Faculté des Sciences, B.P. 8106,
Agadir, Maroc.

Abstract: Blue mold, caused by Penicillium italicum, is one the most economically important
postharvest disease of citrus fruits. The control of this mold is largely dependent on application of
synthetic fungicides, which has adverse effects on the environment and promotes the development of
strains resistant to the active components used. To overcome this problem taking into consideration
the increased concern of consumers towards pesticide use and the increasing demand in the quality of
products, alternative methods are needed. Biological control using naturally-occurring substances is a
promising alternative that has been recently explored for managing postharvest decay of fruits. The
screening of antifungal activity of aqueous plants extracts against P. italicum was evaluated using the
agar plates method. Our results show that from a total of 45 aromatic and medicinal plants assessed,
23 reduced the incidence of disease to less than 50%. From which two species, belonging respectively
to the family of Lamiaceae and Asteraceae, completely inhibited the mycelial growth of P. italicum.

Key words: Citrus, Penicillium italicum, postharvest, biological control, antifungal activity, medicinal
and aromatic plants

Introduction

Citrus is the world’s first fruit crop grown in over 100 countries (Smilanick et al., 2005), it is
also one of the key element in Morocco’s crop exportations. More than 65 % of these
exportations are yielded in Souss-Massa-Draa, valley. Although, Citrus culture is subjected to
many postharvest diseases, particularly green and blue mold caused by Penicilium digitatum
and Penicillium italicum respectively, that reduce citrus fruit’s shelf life and that are
subsequently responsible of 20 to 30% of fruit losses (Zhu et al., 2006).
Currently, measures employed to manage both disease involve the application of
fungicides usually incorporated into waxes (Smilanick & Sorenson, 2001). However, the use
of fungicides is becoming increasingly restricted due to stringent regulation, pathogen
resistance development and public concern about chemical residues in fruit and their products
(Jiuxu & Patricia, 2003). Therefore, alternative methods for decay control need to be
developed. Various alternative measures such as the application of food additives,
microorganisms or plant extracts in combination or in a replacement for fungicide have been
developed in order to reduce the input of fungicides. In this study, the antifungal capacities of
the aqueous extracts derived from 45 medicinal and aromatic plants used in southern
Moroccan traditional medicine against P. italicum were investigated.

51
52

Material and methods

Plant material
A total of 45 plants samples were collected in various locations in Souss-Massa-Draa valley
Agadir, Morocco. Different parts of plant were used (stem, leaves, flowers, fruits and seeds).
Tests were carried out with plant powder by the agar plate method (Table 1).

Pathogen culture
Penicillium italicum was isolated from decayed lemon fruit stored at 5°C. The pathogen was
maintained in Potato Dextrose Agar (PDA) and stored at 5°C until use.

Aqueous extract preparation

The plant samples were first air-dried and ground. The powder of each sample was added to
PDA medium, stirred for 10min and then autoclaved for 20min at 121°C. The resulting
suspension was filtered through 4 layers of cheese cloth before being dispensed into 9cm Petri
dishes (Ameziane et al., 2007).
Agar plates were inoculated with P. italicum using a 5mm diameter disk cat from
actively growing edge of one week-old culture and incubated at 24 ± 1°C for 8 days.
The antifungal activity of these extracts was expressed in term of the percentage of
mycelial growth inhibition and calculated according to the formula below:
% inhibition = dc – dt/dc x 100
Where dc = Average increase in mycelial growth in control sets, dt = Average increase in
mycelial growth in treatment sets after 8 days of incubation (Singh & Tripathi, 1999). The
antifungal effect was measured under a totally random design with three replications.

Results and discussion

Forty-five aqueous extracts were assayed for their ability to suppress the growth of P.italicum
in vitro. The distribution of the assessed species according to their botanical families and rate
of protection were summarized in Table1.
The results of the screening were shown in Figure 1. Among 45 plants screened, 23
aqueous extracts have recorded significant antifungal activity. The percents of inhibition of
these extracts were more than 50%, from which two species belonging respectively to the
family of Lamiaceae and Asteraceae completely inhibited the mycelial growth of P. italicum.
Ameziane et al. (2007) found that aqueous extract from Thymus leptobotris, a plant belonging
to the family of Lamiaceae completely inhibited the mycelial growth of Penicillium italicum,
Penicillium digitatum and Geotricum candidum, the main postharvest fungi in citrus crop.
The remaining aqueous extracts showed either reduced inhibitory effect (less than 50%)
or enhanced the mycelial growth of the studied fungus.
Other studies demonstrated the antifungal effect of plant extracts and essential oils and
encouraged the use of these naturally occurring products as an alternative to chemical
fungicides to control postharvest pathogens (Amiri et al., 2008; Tripathi et al., 2008 and
Satish et al., 2007).
 53

Table 1: Distribution of the assessed species according to their botanical families and rate of
growth inhibition

Botanical families Species per family Growth inhibition (%) per species
I< 50 50 ≤ I < 80 80 ≤ I ≤100
Zygophyllaceae 2 1 1
Capparaceae 1 1
Asclepiadaceae 1 1
Lamiaceae 13 10 2 1
Solanaceae 2 2
Marsileaceae 1 1
Anacardiaceae 2 1 1
Rosaceae 3 1 2
Rutaceae 2 1 1
Asteraceae 9 1 3 5
Boraginaceae 1 1
Cistaceae 4 1 2 1
Fabaceae 1 1
Composeae 1 1
Ombilifereae 1 1
Chenopodiaceae 1 1
Total 16 45 45

Figure 1: Effects of water extracts of aromatic and medicinal plants on the growth of
Penicillium italicum
 54

Conclusion

In the present work we demonstrate that several Moroccan medicinal and aromatic plants
possess a potent antifungal activity against P. italicum. Fifty-one percent of the assessed
plants reduced the in vitro growth of P. italicum to less than 50%, from which 39% exhibited
higher inhibition percentages ranging from 80% to 100%. Two species belonging respectively
to the family of Lamiaceae and Asteraceae, completely inhibited the mycelial growth of
P. italicum. Further phytochemical research is needed to identify the active principle for the
antifungal effects of each plant.

References

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