In Vitro Inhibitory Effect of Some Plant Extracts Against Fusarium culmorum (W.G. Smith) Causal Agent of Foot and Root Rot Disease on Wheat

Abstract

Investigations on alternative control methods to chemicals including usage of plant extract for plant
fungal disease take important place in current researches. Plant extracts are generally deemed to be less
hazardous than synthetic compounds and could be alternative to antifungal treatments.
In this study, the purpose was screening of some plant extracts against Fusarium culmorum causing foot
and root rot on different small-grain cereals, in particular wheat and barley. In total 6 plant species including
levander (Lavandula angustifolia), common mullein (Verbascum thapsus), common sorrel (Rumex acetosa),
wood avens (Geum urbanum), poison hemlock (Conium maculatum), henbane (Hyoscyamus niger) consisted
the materials in the study. The antifungal effects of three different concentrations (20%, 40%, and 80%) of
each extracted plant, obtained from dried plant parts in methanol, were evaluated on growth of F. culmorum
by dual test technic on PDA (Potato Dextrose Agar).
The maximum in vitro inhibitory efficiencies were obtained from common mullein (Verbascum
thapsus) at concentrations of 80% and 40% with the ratios of 46.66% and 33.33% respectively, while no
repressive activity exhibited by other plant extracts on pathogen growth on PDA.

Full text

IN VITRO INHIBITORY EFFECT OF SOME PLANT EXTRACTS AGAINST
Fusarium culmorum (W.G. Smith) CAUSAL AGENT OF FOOT AND ROOT ROT
DISEASE ON WHEAT*
Gül İmriz 1
, Fatih Özdemir1, Murat Nadi Taş1, Eyüp Başer1, İlker Topal1, Birol
Ercan1, Mehmet Sait Karaca1
1Bahri Dagdas International Agricultural Research Institute, Konya/Turkey
Corresponding author’s e-mail: gulimriz@hotmail.co.uk
ABSTRACT
Investigations on alternative control methods to chemicals including usage of plant
extract for plant fungal disease take important place in current researches. Plant extracts
are generally deemed to be less hazardous than synthetic compounds and could be
alternative antifungal treatments.
In this study, the purpose was screening of some plant extracts against Fusarium
culmorum causing foot and root rot on different small-grain cereals, in particular wheat
and barley. In total 6 plant species including levander (Lavandula angustifolia),
common mullein (Verbascum thapsus), common sorrel (Rumex acetosa), wood avens
(Geum urbanum), poison hemlock (Conium maculatum), henbane (Hyoscyamus niger)
consisted the materials in the study. The antifungal effects of three different
concentrations (20%, 40%, and 80%) of each extracted plant, obtained from dried plant
parts in methanol, were evaluated on growth of F. culmorum by dual test technic on
PDA (Potato Dextrose Agar).
The maximum in vitro inhibitory efficiencies were obtained from common mullein
(Verbascum thapsus) at concentrations of 80% and 40% with the ratios of 46.66% and
33.33% respectively, while no repressive activity exhibited by other plant extracts on
pathogen growth on PDA.
Keywords: plant extracts, Fusarium culmorum, in vitro, fungistatic activity
BUĞDAYDA KÖK VE KÖK BOĞAZI ÇÜRÜKLÜK ETMENİ Fusarium
culmorum (W.G. Smith)’ A KARŞI BAZI BİTKİ EKSTRAKTLARININ IN
VITRO İNHİBİTÖR ETKİLERİ
ÖZET
Son yıllarda, bitki fungal hastalıklarına karşı kimyasallara alternatif mücadele
yöntemleri üzerine yapılan araştırmalarda bitki ekstraktları önemli yer tutmaktadır. Bitki
ekstraktlarının sentetik bileşiklere göre daha az zararlı olduğu bilinmekte ve potansiyel
bir antifungal tedavi yöntemi olarak değerlendirilmektedir.
Çalışmanın amacı, başta buğday ve arpa olmak üzere küçük-daneli tahılların kök ve kök
boğazı çürüklük etmeni Fusarium culmorum’a karşı bazı bitki ekstraktlarının
engelleyici etkilerinin belirlenmesidir. Çalışmanın materyali olarak Lavanta (Lavandula
angustifolia), sığırkuyruğu (Verbascum thapsus), labada (Rumex acetosa), meryem otu
*The article was presented as a poster at 2nd ICSAE (International Conference on Sustainable Agriculture
and Environment)

(Geum urbanum), baldıran (Conium maculatum), banotu (Hyoscyamus niger) olmak
üzere toplam 6 bitki türü kullanılmıştır. Kurutulmuş bitki parçalarından metanolde
ektraktlar elde edilmiş ve her bitki ekstraktının 3 farklı konsantrasyonunun (%20, %40
ve %80) F. culmorum’un PDA üzerinde ki gelişmine etkisi ikili kültür tekniği ile
belirlenmiştir.
En yüksek inhibitor etkisi sığırkuyruğu ekstraktının %80 ve %40 oranındaki
konsantrasyonlarından elde edilirken, diğer ekstraktların hiçbirisi PDA üzerinde
fungusun gelişimi üzerinde engelleyici bir etki göstermemiştir.
Anahtar kelimeler: bitki ekstraktları, Fusarium culmorum, in vitro, fungistatik aktivite
INTRODUCTION
Wheat production is affected by many of plant pathogenic organisms including fungi.
Fusarium culmorum is a ubiquitous soil-borne fungus able to cause foot and root rot on
different small-grain cereals, in particular wheat and barley. It causes significant yield
and quality losses. Foot and root rot pathogen Fusarium culmorum (W.G. Smith) Sacc.
also known as Fusarium crown rot causes some typical symptoms such a seedling blight
with death of the plant before or after emergence, brown discoloration on roots and
coleoptiles of the infected seedlings, brown discoloration on sub-crown internodes and
on the first two/three internodes of the main stem, tiller abortion, formation of
whiteheads with shriveled white grains. For controlling such plant diseases, application
of fungicides is inadequate in some cases besides their hazardous effects for
environment and living organisms.
Growth of fungal pathogens has led to considerable economic losses in agricultural
crop. As a solution, synthetic fungicides have been used globally since the 1950s to
protect major crops from damage by phytopathogenic fungi (Knight et al., 1997; Leroux
et al., 2010). As an alternative solution to chemicals, plant extracts are generally deemed
to be less hazardous than synthetic compounds and could be appropriate antifungal
treatments. Antifungal activity of some isolated principles from plant extracts may be
more effective than some commercial synthetic fungicides. The presence of naturally
occurring substances in plants with anti-microbial properties have been recognized and
tested against a wide range of pathogenic microbes (Tamuli et al., 2104). With the
increase of interest in antibiotics plants as a source of potential antimicrobial substances
are receiving considerable attention throughout the world. Recently many aqueous plant
extracts have been shown to have inhibitory action against some plant as well as human
pathogenic microbes. Nowadays some synthetic as well as semi-synthetic antimicrobial
agents have been developing, among which very few have broad spectrum activity and
most of them are environmentally hazardous in nature. The extensive use of
agrochemicals especially fungicides, resulted more carcinogenic risk than other
pesticides which may give rise to undesirable biological effects on animals and human
beings (Osman and Abdulrahman, 2003).
Many reports are available on the effects of plants on fungal growth and mycotoxin
production: thyme, sage, origano, coriander (Yıldız et al., 2001; Yanar, 2014; Mirik and
Aysan, 2005), clove (Hitokoto et al., 1980), cinnamon, rosemary, lavender (Bishop and
Thornton, 1997; Erdoğan et al., 2014), cumin, pepper (Abdou et al., 1972), garlic, onion
*The article was presented as a poster at 2nd ICSAE (International Conference on Sustainable Agriculture
and Environment)

(Yıldız et al., 2001; Gandhi and Ghodekar, 1988), basil, saffron, marjoram and anise
(Hitokoto et al., 1980), mentha (Erdoğan et al., 2014) are examples. In this study, it was
aimed to assess in vitro inhibitor activities of 6 plant species including levander
(Lavandula angustifolia), common mullein (Verbascum thapsus), common sorrel
(Rumex acetosa), wood avens (Geum urbanum), poison hemlock (Conium maculatum),
henbane (Hyoscyamus niger), eucalyptus against foot and root rot pathogen Fusarium
culmorum.
MATERIAL and METHOD
Plant materials and pathogen
The leaves, seeds and stem parts of selected plant species including levander
(Lavandula angustifolia), common mullein (Verbascum thapsus), common sorrel
(Rumex acetosa), wood avens (Geum urbanum), poison hemlock (Conium maculatum),
henbane (Hyoscyamus niger) collected from Konya-Turkey, dried at room temperature
(25ºC-30ºC). The pathogen isolate was obtained from of Prof. Dr. Berna Tunalı (19
Mayıs University, Samsun)’s culture collection.
Preparation of the extracts from plant materials
The extraction of plant materials was done according to the methods defined by Tavares
et al. (2009). The dried plant parts were grounded into powder form and packaged
stored in polyethylene bags at room temperature until needed. 50 g of each powdered
plant sample was weighed and added into a bottle containing 500 ml of methanol
(Merck % 99.5). The bottles were closed tightly and incubated at room temperature for
7 days. During incubation period, the bottles were shaken twice a day. After incubation
period, each suspension was filtered individually (Whatman Filter Paper No:1), and
extracted liquids was transferred to covered glass bottles. The methanol of these
extractions was evaporated by vacuum Rotary Evaporator (Heidolphe-VAP Precision) at
42±2°C. The extractions were kept in water bath at 42ºC for a day to evaporate the
methanol completely. The pure plant extracts then filter sterilized and were transferred
to sterile flacon tubes in amber color to avoid from light and kept at +4ºC to use when
needed. The extracts were diluted with methanol (99.5%) at desired volume (%v/v).
In vitro inhibitory activities of plant extracts on Fusarium culmorum
Three concentrations (20%, 40%, and 80%, v/v) of each extract were included in the
study to determine their inhibitor efficiencies against F. culmorum growth on PDA. In
order to have pathogen inoculum, F. culmorum was grown on PDA for a week.
In order to determine the efficiency of extracts, dual test technique was applied (Ulke,
2003). A sterilized filter paper in 0.7 cm diameter was placed on PDA petri dishes as 3
cm from the center and 50µl of extract was soaked. A pathogen disc freshly grown (7
days old) on PDA was placed on opposite of the extract disc as 3 cm from the center in
the same petri dish. The petri dishes were sealed with Parafilm. The experiments were
carried out in 3 replicates per treatment. As control of the experiment, only sterile
distilled water was used instead of extract treatment. Fungal growth was observed and
*The article was presented as a poster at 2nd ICSAE (International Conference on Sustainable Agriculture
and Environment)

recorded after 7 days incubation at 25±1ºC. Inhibition percentage of fungal growth was
calculated by using the following formula (Bhuyan et al., 2015);
%Inhibition=100
(
Control−Treatment
)
Control
RESULTS and DISCUSSION
As seen in Table 1, among the extracts the best inhibitory effects (46.66% and 33.33%)
were obtained from common mullein (Verbascum thapsus) at concentrations of 80% and
40% (v/v) respectively (Figure 1) while no efficiency was observed at concentration of
20% (v/v). It was observed that inhibition percentage of the extract increased with the
increase in concentration (Table 1). Besides, none of other extracts did exhibit any in
vitro inhibitory activity against growth of F. culmorum on PDA.
Figure 1. The in vitro efficacies of common mullein extracts at of 80% (left) and 40%
(right) (v/v) concentration
Table1. The inhibitory effects (inhibition percentage) of the extracts on growth of
Fusarium culmorum on 7th days of inoculation
Plants extracts Extract
Concentration
v/v (%)
Means of
Inhibition
Zones (cm)
Inhibitory
Efficiency
(%)
Common mullein
(Verbascum thapsus)
80%
40%
20%
4.2
3.0
0.0
46.66
33.33
0.00
Levander
(Lavandula angustifolia)
80%
40%
20%
0.0
0.0
0.0
0.00
0.00
0.00
Common sorrel
(Rumex acetosa)
80%
40%
20%
0.0
0.0
0.0
0.00
0.00
0.00
Wood avens
(Geum urbanum)
80%
40%
20%
0.0
0.0
0.0
0.00
0.00
0.00
Poison hemlock
(Conium maculatum)
80%
40%
20%
0.0
0.0
0.0
0.00
0.00
0.00
Henbane
(Hyoscyamus niger)
80%
40%
20%
0.0
0.0
0.0
0.00
0.00
0.00
*The article was presented as a poster at 2nd ICSAE (International Conference on Sustainable Agriculture
and Environment)

Control
(Sterile distilled water)
- 0.0 0.00
The inhibition of the growth of the pathogenic fungi is due to the active ingredients in
the plant (Shetty et al., 1989). Besides, Lubaina and Murugan (2013), indicated that the
efficacy of medicinal plant extracts may induce a systemic resistance in host plants
against pathogens. The extracts may also retard the development of infections at early
growth stages by inhibiting the mycelial growth of pathogens (Krebs and Forrer, 2001),
or the toxic effect of active principles may act directly on the pathogens (Amadioha,
2000).
In the present study, the common mullein extract showed a fine inhibition on the hyphal
growth of soil-borne pathogenic fungi F. culmorum on PDA with increasing
effectiveness depending on the concentrations. There is a number of pharmacological
activities such as anti-inflammatory, antioxidant, anticancer, antimicrobial, antiviral,
antihepatotoxic and anti-hyperlipidemic activity have been ascribed to this plant (Riaz
et al., 2013). Dulger et al. (2015), investigated the antibacterial potential of the leaves of
common mullein against the pathogens causing complicated urine tract infections.
According to their results, the extract showed strong antimicrobial activity against
Escherichia coli, Enterococcus faecalis and Candida albicans.
Many studies shows the success of common mullein in pharmacology, however, the
number of studies with this plant on plant pathogens seems quite few. The findings
reported by Turker and Camper (2002) stated that Agrobacterium tumefaciens-induced
tumors in potato disc tissue were inhibited by the aforesaid plant extracts. Moreover, the
researchers reported antibacterial activity of common mullein (especially the water
extract) with Klebsiella pneumonia, Staphylococcus aureus, Staphylococcus
epidermidis and Escherichia coli.
The present research are in line with the researches infer that leaf extracts in general
have great potentiality in the control of fungal diseases in commercially important crop
plants. Soil-borne pathogens such as Fusarium culmorum are quite difficult to manage
due to there is no chemical to control the disease strictly. The disease may be managed
economic and eco-friendly by using an effective plant extracts, since the plants are
existing in nature as wild. More detailed studies, however, including characterization
bioactive compounds, effect on plants in vivo and natural conditions, are needed to be
accomplished with the promising plant extracts.
ACKNOWLEDGEMENT
We would like to thank to Prof. Dr. Berna Tunalı for her helpfulness and supplying the
Fusarium culmorum isolate from her culture collection to our study.
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*The article was presented as a poster at 2nd ICSAE (International Conference on Sustainable Agriculture
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