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Original Article 

Biosci. J., Uberlândia, v. 33, n. 3, p. 566-571, May/June. 2017 

EVALUATION OF THE FUNGITOXIC ACTIVITY OF VEGETAL 
EXTRACTS ON THE MYCELIAL GROWTH OF PHYTOPATHOGENS 

 
AVALIAÇÃO DA ATIVIDADE FUNGITÓXICA DE EXTRATOS VEGETAIS SOBRE O 

CRESCIMENTO MICELIAL DE FITOPATÓGENOS 
 

Talita Pereira de Souza FERREIRA1; Gil Rodrigues dos SANTOS1;  
Tatiani Pereira de Souza FERREIRA1; Tallyta Santos TEIXEIRA1; Ryhára Dias BATISTA1; 

Raiana Batista da LUZ1; Luiz Gustavo de Lima GUIMARÃES2 
1. Universidade Federal do Tocantins, Gurupi, TO, Brasil. cupufer@gmail.com; 2. Universidade Federal de São João Del Rei, 

São João Del Rei, MG, Brasil. 
 

ABSTRACT: Although the productivity of common bean in Tocantins is economically favorable, it has been 
infected by various pathogens found in soil. Among the major diseases is the web blight and root rot caused by 
Rhizoctonia solani and collar rot caused by the fungus Sclerotium rolfsii. This study aimed to evaluate the fungitoxic 
activity of methanol extracts of eight plant species on the inhibition of mycelial growth of S. rolfsii and R. solani. The 
fungitoxic activities were carried out over the inhibition of mycelial growth by means in vitro assays. The extracts were 
applied in concentrations of 250, 500, 1000, 2500 e 5000 µg ml-1 in PDA culture medium. In bioassays, it was found the 
significant effect of plant, concentration and also their interaction on the antifungal activity of the extracts. However, some 
extracts showed no inhibition of mycelial growth of the pathogens studied. Among those who had higher inhibitions is the 
extract of Lantana trifolia, which inhibited the mycelial growth of S. rolfsii in all concentrations, being the same as 97% 
for the highest concentration. When the methanol extract of Piper amplum Kunth, inhibition of the highest concentration 
was 83% for S. rolfsii and 74% for R. solani. These results show the potential of methanolic extract of Lantana trifolia and 
Piper amplum Kunth in the control set of plant pathogens studied.  
 

KEYWORDS: Methanol extract. Sclerotium rolfsii. Rhizoctonia solani. Phaseolus vulgaris. 
 

INTRODUCTION  
 

The common bean (Phaseolus vulgaris) is 
the most important legume crop consumed by 
humans worldwide. Various pathogens of 
epidemiological importance can damage common 
bean crops reducing their quality (PARSA et al., 
2016). The risk of losses caused by pests and 
diseases in bean decreases the profitability for 
producers, resulting in decreased exploitation of this 
legume by large producers (GONÇALVES-
VIDIGAL et al., 2007; CUNHA et al., 2005). 
Therefore, there is a growing interest in discovering 
plant-derived anti-microbial substances as 
alternative agents   against fungal growth (EL-
MOUGY et al., 2007). 

Of the 60 diseases that can affect bean 
crops, 31 are caused by fungi. These pathogens are 
reported as the primary causative agents of diseases 
in the economically important vegetable crops. Each 
year they destroy a third of agricultural production 
worldwide resulting in approximately 20% 
reduction in yield of cultivars (GARCIA et al., 
2002). Among the diseases caused by pathogenic 
fungi in  bean crops, there are the collar rot and the 
web blight which are caused by the fungi Sclerotium 
rolfsii  and  Rhizoctonia solani, respectively. 

So, to avoid significant losses caused by 
diseased plants, several control methods are 
currently employed. The most common being the 
use of synthetic fungicides (BAJWA; KHALID; 
CHEEMA, 2003). However, the abusive use of 
these compounds has been causing environmental 
damage and has even been proven harmful for 
consumers, besides promoting  resistance of the 
pathogenic microorganisms. Furthermore, these 
synthetic fungicides also affect the symbiotic 
microbial population present in the local ecosystem 
(CUNICO et al., 2004). In view of this, ongoing 
research is being conducted in order to create and/or 
discover new methods to halt the advance of 
pathogenic fungi growth which are environmentally 
friendly and effective in the action. 

Extracts from different parts of medicinal 
plants have been the source of many compounds 
that have antimicrobial activities. Many studies have 
reported the fungitoxic activity of various plant 
extracts. For instance, Vogt et al. (2012), while 
studying the effects of the extract of a medicinal 
plant from Argentina, Larrea divaricata, 
corroborated its empirical use in the treatment of 
some plant diseases and showed high rate of 
antifungal activity, making it a potential natural 
source for biological control. Fiori et al. (2000) 
reported that the extracts from the leaves of 

Received: 13/04/16 
Accepted: 20/11/16 



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Evaluation of the fungitoxic…       FERREIRA, T. P. S. et al 

Biosci. J., Uberlândia, v. 33, n. 3, p. 566-571, May/June. 2017 

Eucalyptus citriodora inhibited the mycelial growth 
of the pathogenic fungus Didymella bryoniae, while 
the plant extract of Achillea millefolium inhibited 
the germination of spores of the same fungus. Also 
in this work, the plant extract Ageratum conyzoides 
both inhibited the growth of mycelial and spore 
germination of the fungus Didymella bryonae. 

Given the above, this work aimed to study 
the fungitoxic activity of methanol extracts of eight 
plant species on the inhibition of mycelial growth 
Sclerotium rolfsii  and Rhizoctonia solani, in order 
to develop alternative and effective means to control 
the collar rot, web blight and root rot caused by 
these pathogens in the culture of the common bean. 
 
MATERIAL AND METHODS 
 
Collection and extraction of vegetal plant 
extracts 

Eight plant species were studied: ‘sucupira’ 
(Pterodon emarginatus), ‘mastruz’ (Chenopodium 
ambrosioides), mint (Mentha piperita), terramycin 
(Alternanthera dentata), ‘negramina’ (Siparuna 
guianensis Aublet), pepper (Piper amplum Kunth), 
‘camará’ (Lantana trifolia L.) and  ‘grandiúva-
d'anta’ (Psychotria leiocarpa Cham. Et 
Schlechtend). After harvesting, the plant material 
was sent to the Chemistry Laboratory on the UFT 
Campus in Gurupi, where they underwent drying 
and processes to obtain the extracts. The leaves 
were dried in the shade at room temperature and 
subsequently were cut and submitted to extraction 
with cold methanol. Thirty grams of dried leaves 
were used with 1.5 liters of solvent for each 
extraction for a period of 7 days. After this period, 
the mixture was filtered and evaporated under 
reduced pressure to obtain the extracts (COSTA et 
al., 2008).  
 
Isolation of fungi used in biological assay 

The pathogens S. rolfsii and R. solani were 
isolated from plants of the common bean Phaseolus 
vulgaris with symptoms of the disease (collar rot 
and root rot) as follows: plant parts with symptoms 
of the disease were cut and washed in water. Then 
these pieces were surface sterilized by successive 
immersion in 70% ethanol for 30 seconds, a solution 
of 1% sodium hypochlorite for 40 seconds and three 
times in demineralized water. After being washed 
several times, they were transferred with sterile 
tweezers to a Petri plate containing 15 ml of PDA 
culture medium (Potato Dextrose Agar) and 1 mg of 
ampicillin. Daily, the Petri dishes were monitored 
and fungal colonies that showed no contaminants 

were transplanted into new plates with the same 
culture medium (VALADARES et al. 2008). 
 
Experimental design and biological activity 

Isolated pure cultures of plant pathogens S. 
rolfsii and R. solani that were 7 days old were 
separately tested to evaluate fungitoxicity. 
Bioassays were conducted by adding to the PDA 
culture medium (Sigma-Aldrich®), aqueous 
solutions of methanol extracts from the eight 
lyophilized medicinal plants, examined in 
appropriate volumes to obtain concentrations of 
250, 500, 1000, 2500 and 5000 µg mL-1, equally for 
each microorganism. With the medium already 
poured into Petri dishes, discs of agar-mycelium of 
5 mm diameter made of pure cultures of fungi were 
inoculated on the surface of each culture with their 
respective treatments. Subsequently, the plates were 
sealed with plastic film and incubated in a growth 
chamber (25 ± 1 °C) and exposed to a photoperiod 
of 12 hours. To estimate the efficiency of the 
treatments, the diameter of each of the colonies was 
measured during the mycelial growth and compared 
to that of absolute witness. It was observed that it 
completely covered the surface of the medium. The 
radial growth was measured with calipers in two 
orthogonal axes to each other by calculating an 
average for each plate. The experimental design was 
completely randomized with three replications. 
Treatments were arranged in a factorial scheme 2 x 
8 x 5 for evaluation as antifungal activity being 
constituted by combinations of the eight plant 
species, two pathogenic fungi and concentrations of 
extracts. Analysis of variance was carried out to 
check the effects of treatments, using regression 
analysis to assess the percentage of inhibition of 
mycelial growth of phytopathogenic fungi, in 
relation to concentration using the statistical 
program SISVAR. 
 
RESULTS AND DISCUSSION 
 

According to the results of the assessment 
of the fungitoxic activity of plant extracts from eight 
plant species studied, there was a significant effect 
on plant factors, concentration, and also the 
interaction of these factors on antifungal activities 
presented by the extracts and applied to each test 
performed. Through this interaction, we observed 
the existence of a variation of antifungal activity, 
depending on the extract of each plant and the 
concentration of each. The values of antifungal 
activity in the plant extracts in relation to their 
concentrations are assessed by inhibition assay in 



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Evaluation of the fungitoxic…       FERREIRA, T. P. S. et al 

Biosci. J., Uberlândia, v. 33, n. 3, p. 566-571, May/June. 2017 

percentages for the two fungi S. rolfsii and R. solani are shown in Figure 1. 

 
Figure 1. Fungitoxic activity of the methanol extracts of the ten plant species on mycelial growth of pathogenic fungi 

Sclerotium rolfsii and Rhizoctonia solani. A: Piper amplum; B: Lantana trifolia; C: Mentha piperita; D: 
Chenopodium ambrosioides; E: Psychotria leiocarpa; G: Siparuna guianensis; H: Pterodon emarginatus; I: 
Alternanthera dentata. 



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Biosci. J., Uberlândia, v. 33, n. 3, p. 566-571, May/June. 2017 

The effect of the extract L. trifolia (Figure 
1B) against R. solani was observed when the 
witness completely covered the Petri dish. The 
mycelial growth was reduced at concentrations of 
1000, 2500 and 5000 µg ml-1, and there was 
inhibition at the highest concentration of 64%. 
Already with the fungus, S. rolfsii, the methanol 
extract showed this inhibitory effect at all 
concentrations used. An inhibition of 97% was 
observed in the highest concentration. 

Likewise, the extract of Psychotria 
leiocarpa (Figure 1E) significantly reduced the 
growth of mycelia of both fungi studied. The 
inhibition was verified from concentration of 5000 
µg ml-1, while the highest concentration obtained 
72% mycelial inhibition in S. rolfsii and 55% in R. 
solani. When these same fungi were exposed to the 
extract Piper amplum (Figure 1A), inhibition of the 
highest concentration was 83% for S. rolfsii and 
75% for R. solani, considering the inhibitions were 
observed from lower concentrations. 

According to the regression analyses for the 
two pathogenic fungi, mycelial inhibition in relation 
to concentrations of methanol extracts of L. trifolia 
(Figure 1B) and of P. leiocarpa (Figure 1E), which 
follow equations of the second order. Already 
having considered the concentrations of the extract 
of P. amplum (Figure 1A), which follow equations 
of the first order. 

The assessment of the potential fungitoxic 
plant extract A. dentata (Figure 1H), showed no 
effect on the inhibition of mycelial pathogen S. 
rolfsii. On the other hand, the pathogen R. solani 
was sensitive to all extracts of plants assessed. But 
some extracts even at the highest concentration 
showed lower percentage inhibition of mycelial 
growth even though the extracts; P. emarginatus 
(Figure 1G), C. ambrosioides (Figure 1D), S. 
guianensis (Figure 1F) and A. dentata (Fig. 1H), 
exhibited the following activities: 24, 24, 14 and 
20%, respectively. Considering the fact that 
concentration varies according to the regression 
analyses, the mycelial inhibition of these extracts 
followed equations of the second degree, except for 
the inhibitions caused by the different 
concentrations of the extract A. dentata (Figure 1H), 
which followed an equation of first degree. The 
extracts of M. piperita (Figure 1C), C. ambrosioides 
(Figure 1D), S. guianensis (Figure 1F) and P. 
emarginatus (Figure 1G) also had no significant 
effects on the inhibition of mycelial growth of 
phytopathogenic fungus S. rolfsii, since before their 
concentration was increased, the mycelial growth of 
this microorganism was inhibited by 36, 4, 6 and 
10%, respectively. However, the methanol extract of 

M. piperita (Figure 1C) on R. solani showed a 
significant inhibition of 51% at its highest 
concentration. 

Many studies have presented results which 
corroborate those found in this work, as conducted 
with plant extracts of Ocimum basilicum (basil), 
Ruta graveolens (rue), Baccharis trimera (gorse), 
which showed antifungal activity on Rhizoctonia 
solani, Sclerotium rolfsii, Alternaria alternata, 
Phythophtora sp. and Colletotrichum graminicola 
(STARGALIN et al., 1999). Significant effects were 
observed in a concentration of 1% applied to the 
turmeric extract which has led to a greater than 50% 
inhibition of growth of Fusarium oxysporum (61%) 
and R. solani (61%) (AMARAL; BARA, 2005). 
Thangavelu et al. (2004) showed that extracts of 
Solanum torvum, Jatropha glandulifera and 
Emblica officinalis were highly inhibitory to 
mycelial growth of Colletotrichum musae, such 
inhibitions being directly related to the 
concentration factor. Extracts of leaves and seeds of 
Pithecellobium dulce had fungitoxic effects in more 
than 50% mycelial growth and sporulation of 
Botrytis cinerea, Penicillium digitatum and 
Rhizopus stolonifer, the strawberry fruit 
(BAUTISTA-BAÑOS et al., 2003).  
 
CONCLUSIONS 

 
Among the methanol extracts of the plants 

studied, the one showing the best antifungal effect 
on mycelial growth of the fungus S. Rolfsii was the 
methanol extract of the plant L. trifolia with 
inhibition of 97%.  

For R. Solani, the methanol extract of P. 
amplum showed the best result among all extracts 
evaluated, inhibiting 75% growth of mycelia. 
Furthermore, observing the percentages of inhibition 
of the two fungi in general, S. rolfsii was more 
resistant to plant methanol extracts than all 
pathogens studied.  

These results show that the potential of 
methanol extracts for L. trifolia and P. amplum in 
the control group of plant pathogens studied. 
However, it is necessary to conduct further studies 
employing fractionation on the extract, for 
identification of molecule(s) performing such 
activities. 
 
ACKNOWLEDGEMENTS 
 

This work was supported by the Conselho 
Nacional de Desenvolvimento Científico e 
Tecnológico – CNPq – Brazil. 



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Biosci. J., Uberlândia, v. 33, n. 3, p. 566-571, May/June. 2017 

 
RESUMO: Apesar da produtividade do feijão comum no Tocantins ser economicamente favorável, o mesmo 

pode ser infectado por vários patógenos habitantes do solo, dentre as principais doenças encontra-se a mela e a podridão 
radicular causadas pelo fungo Rhizoctonia solani e a podridão do colo causada pelo fungo Sclerotium rolfsii. Este trabalho 
teve como objetivo avaliar a atividade fungitóxica dos extratos metanólicos de oito espécies vegetais sobre a inibição do 
crescimento micelial de Sclerotium rolfsii e Rhizoctonia solani. As atividades fungitóxicas foram realizadas perante a 
inibição do crescimento micelial por meio de ensaios in vitro, sendo os extratos aplicados nas concentrações de 250, 500, 
1000, 2500 e 5000 µ g ml-1 em meio de cultura BDA. Observou-se o efeito significativo dos fatores planta, concentração e 
também da interação destes sobre as atividades fungitóxicas. No entanto, alguns extratos não apresentaram inibição do 
crescimento micelial dos fitopatógenos estudados. Entre os que apresentaram maiores inibições encontra-se o extrato de 
Lantana trifolia, que inibiu o crescimento micelial do S. rolfsii em todas as concentrações, sendo o mesmo de 97% para a 
maior concentração. Já a concentração mais elevada do extrato metanólico de Piper amplum apresentou inibição de 83% 
sobre o crescimento micelial de S. rolfsii e 74% sobre o crescimento micelial de R. solani. Tais resultados evidenciam a 
potencialidade dos extratos metanólicos das folhas de Lantana trifolia e de Piper amplum no controle dos fitopatógenos 
estudados.  
 

PALAVRAS-CHAVE: Extrato metanólico: Sclerotium rolfsii. Rhizoctonia solani. Atividade fungitóxica. 
 
 
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