Volume: 3: Issue-1: Jan - Mar-2012

ISSN 0976-4550

Accepted: Nov-2011

Research Article

SCREENING OF ANTIFUNGAL EFFECTS OF PSEUDOCLITOCYBE

CYATHIFORMIS Bull. (Singer)
Perihan Gler1*, Fatih Kutluer2,Takn Erol3, Erkan Eren4, lknur Kunduz1 and Hayriye Bier1
1*

Corresponding author: Krkkale University Faculty of Science and Arts, Department of

Biology, Yahsihan-Krkkale,Turkey
2
Krkkale University Krkkale Vocational High School, Mushroom Programme, YahsihanKrkkale,Turkey
3
Krkkale University Krkkale Vocational High School, Organic Agriculture Programme,
Yahsihan-Krkkale,Turkey
4
Ege University Bergama Vocational High School, Mushroom Programme, zmir-Turkey

ABSTRACT : In this study, antifungal activities of Pseudoclitocybe cyathiformis extracts with the
help of acetone and chloroform against to Fusarium species (Fusarium culmorum and Fusarium
moniliforme) were investigated.
Pseudoclitocybe cyathiformis was dried at aseptic conditions and put thru extractions for 12 hours in
solvents. Than the evaporator at 40C and finally dried material stored at + 4C.(Jonathan and Fasidi,
2003). Antifungal activities were measured by Disc Diffusion method (Stoke and Ridgway, 1980).
According to this method; the inoculums containing Fusarium culmorum and Fusarium moniliforme
were spread agar medium.
In result antagonistic effects of Pseudoclitocybe cyathiformis was found against both Fusarium spp..
The clear zone of inhibition was observed around the mushroom extracts. For control water and only
acetone and chloroform saturated discs were used.
The results were comprised with commercial antibiotics (amoxycillin and erythromycin).
Key Words: Pseudoclitocybe cyathiformis, Fusarium culmorum, Fusarium moniliforme,
antifungal activity.

INTRODUCTION
Mushrooms are claimed to exhibit antitumour, antiviral, antibiotic, anti-inflammatory, hypoglycaemic,
hypocholesterolemic and hypotensive activities, as well as having therapeutic and nutritional value
(Chang and Miles, 1989; Breene, 1990; Miles and Chang, 1997). Their antitumour activity is the most
thoroughly researched medicinal effect especially that of the shiitake mushroom, the maitake
mushroom, Sclerotina sclerotiorum and Schizophyllum commune (Borchers, 1999). Antibacterial
activity of the methanol extract of three polypore macrofungi Phellinus rimosus, Ganoderma lucidum
and Navesporus floccosa were studied (Sheena et al., 2003).
Studies on antimicrobial properties of Ganoderma lucidum, Fomes lignosus, Schizophyllum commune,
Pleurotus florida, Lentinus subnudus, Leptoporus sp, Panus fulvus, Coriolus versicolor, Trametes
saepiara, Trametes betulina, Daedalea elegans and Auricularia auricula were carried out (Fagedev,
Oyalede, (2009).
The extracts of ethyl acetat, chloroform, aceton and ethyl alcohol of Pleurotus ostreatus (Jack ex Fr.)
Kum var. salignus, Pleurotus florida Fovose, Schizophyllum commune Fr., Helvella leucomelaena
(Pexs) Nannf. and Amanita virosa (Fr.) Bertillon were investigated in respect to their antimicrobial
activities against Escherichia coli ATCC 25922, Staphylococcus aureus and Pseudomonas aureginosa
ATCC 27857 by Diffusion Method (Demirhan et al., 2007).
In this study, antifungal activities of Pseudoclitocybe cyathiformis Bull. (Singer) extracts with the help
of acetone, chloroform against to Fusarium moniliforme and Fusarium culmorum were investigated.

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Perihan Gler et al

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MATERIALS AND METHODS

Organism
Pseudoclitocybe cyathiformis was collected from Trkmenbabada-Eskiehir-Turkey and these
mushrooms were collected and identified by lknur Kunduz and all specimens have been deposited at
the Krkkale University, Turkey.

Test organisms
In this study, Fusarium spp. were used that F.culmorum, F.moniliforme. Test organisms were
obtained Ministry of Agricultural Rural Affairs (MARA)-Turkey. These fungal cultures were
maintained in nutrient broth (Merck).Mycelial agar discs were taken from developed Fusarium spp. on
the potato dextrose agar (PDA) and were incubated in the nutrient broth at 100 rpm for 48 hours and
activated.

Preparation of crude extracts

Pseudoclitocybe cyathiformis were dried at aseptic conditions and were cut into bits. Dried
mushrooms were pulverized in a blender and 50g each of the powdered samples were soaked
separately in 300ml of 95% acetone and chloroform until complete exhaustion in an Erlenmayer flask.
The flasks were covered with aluminum foil and allowed to stand for 7 days for extraction. These
extracts were filtered through Whatman filter paper no.1 and were evaporated in vacuous and dried
using rotary evaporator at 40C. The extracts were collected and dried (Jonathan and Fasidi, 2003).

Statistical analysis
Data were analyzed and treatments compared using the analysis of variance (ANOVA) (p>0.05)

Simple susceptibility screening

The activated test organisms of Fusarium spp. were poured into potato dextrose agar (PDA) separately
as 500l. and were spread with spatula and dried as aseptic. They were settled inside the prepared
acetone and chloroform 6mm extracts for 10 seconds and than left into the Fusarium inoculated agar
medium. The antifungal effects of the extracts were determined disc diffusion method (Stoke and
Ridgway, 1980). They were incubated at 28C in order for their development can be monitored.
At the end of the incubation the sizes of the formed inhibition zones were measured as milimetric and
their photos were taken. Also the acetone and chloroform sucked discs which were used as solvents in
the study tried in the Fusarium inhibition and comparisons are made. The sterile distilled water used in
the dilution of solid mushrooms extracts were used as the control. All the tests were carried out in
triplicates.

RESULTS AND DISCUSSION

In this study; antagonistic effects of Pseudoclitocybe cyathiformis was found against Fusarium
culmorum and Fusarium moniliforme. This was obtained by the clear zone of inhibition produced by
the fungi around the tested mushroom extracts (Figure 1, Figure 2)

1
2

2
1

Figure 1. Antifungal activity of Pseudoclitocybe cyathiformis against F.culmorum

1-Acetone, 2-Chloroform

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Perihan Gler et al

ISSN 0976-4550

Figure 2. Antifungal activity of Pseudoclitocybe cyathiformis against F.moniliforme

1-Acetone, 2-Chloroform
All of antifungal activities of Pseudoclitocybe cyathiformis were shown at Table 1.

Table 1. Antifungal activity of Pseudoclitocybe cyathiformis

Microorganisms
Fusarium culmorum
Fusarium moniliforme

Inhibition zone diameter (mm)

Solvents
Control*
1
2
0
38
30
0
34
36

Control*=Distilled Water; 1-Acetone; 2-Chloroform

As can be seen in Table 1 P.cyathiformis has shown the most activity towards F.culmorum in acetone.
The values of acetone and choloroform as 36 mm and 40 mm (average,38 mm) (p>0.05) and as 27 mm
and 33 mm (average 30 mm) (p>0.05) respectively. F. moniliforme values are in the acetone and
choloroform as 33 mm and 35 mm (average, 34 mm) (p>0.05) and as 33 mm and 39 mm (average 36
mm) (p>0.05) respectively.
Gbolagade and Fasidi (2005) in their study; Auricularia polytricha, Corilopsis occidentalis, Daldinia
concentrica, Daedalea elegans and Tricholoma lobayensis were found to exhibit various degrees of
antagonistic effects against the Aspergillus niger, A.flavus, Candida albicans and Microsporum
boulardii

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