Biota Neotropica 18(1): e20170418, 2018

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ISSN 1676-0611 (online edition)

short communication

Antifungal potential of crude extracts of Trichoderma spp.

Eder Marques1,2, Irene Martins1 & Sueli Correa Marques de Mello1,2,*

1
Empresa Brasileira de Pesquisa Agropecuária, Embrapa Recursos Genéticos e Biotecnologia, Controle Biológico,
Laboratório de Fitopatologia, Brasília, DF, Brazil
2
Universidade de Brasília, Instituto de Biologia, Depto. de Fitopatologia, Brasília, DF, Brazil
*Corresponding author: Sueli Correa Marques de Mello, e-mail: [email protected]

MARQUES, E., MARTINS, I., MELLO, S. C. M. Antifungal potential of crude extracts of Trichoderma spp. Biota
Neotropica. 18(1): e20170418. http://dx.doi.org/10.1590/1676-0611-BN-2017-0418

Abstract: Antibiosis is the mechanism by which certain microorganisms respond to the presence of others, secreting
compounds or metabolites capable of inhibiting or impeding their development. The crude extract of Trichoderma contains
a mixture of secondary compounds, which may show antibiotic effect, and has been used for the prospect of this fungus
for biological control and other industrial purposes. Faced with the increasing demand of agriculture for ecologically
compatible alternatives for the management of diseases, this work aimed to investigate the spectrum of action of Non-
Volatile Metabolites (NVMs) of Trichoderma isolates against different plant pathogenic fungi. The antagonistic potential
of NVMs was evaluated through the incorporation method of the filtered liquid extract in PDA medium. The assays
showed that all the NVMs produced inhibited the fungus Sclerotinia sclerotiorum similarly. On the other hand, strains
CEN1245 and CEN1274, both belonging to the species Trichoderma brevicompactum, showed broad spectrum against
Sclerotium rolfsii, Colletotrichum gloesporioides, Verticillium dahliae, Fusarium oxysporum and Cylindrocladium sp.
The present study describes isolates producing non-volatile metabolites with broad spectrum of antifungal action, as
well as pathogen-specific. The Trichoderma spp. NVMs obtained from different soil samples cultivated with vegetables,
cassava and maize were efficient in inhibiting plant pathogenic fungi belonging to other patossystems, such as forest
or fruit, which could increase their potential application in biological control of plant diseases. In addition, these
antagonistic fungi should be studied in greater depth for the identification of bioactive molecules of industrial interest
or in commercial formulations of products for biological control of plant pathogens.
Keywords: secondary metabolites, antagonism, inhibition of mycelial growth, plant pathogenic fungi.

Potencial antifúngico de extratos brutos de Trichoderma spp.

Resumo: Antibiose é um mecanismo pelo qual certos microrganismos respondem à presença de outros, secretando
compostos ou metabólitos capazes de inibir ou impedir o seu desenvolvimento. O extrato bruto de Trichoderma contém
uma mistura de compostos secundários e tem sido utilizado na prospecção deste fungo para o controle biológico e
demais fins industriais. Diante da crescente demanda da agricultura por alternativas ecologicamente compatíveis para o
manejo de doenças, este trabalho teve como objetivo investigar o espectro de ação de Metabólitos Não Voláteis (MNVs),
produzidos por isolados de Trichoderma, contra diferentes fungos fitopatogênicos. O potencial antagônico dos MNVs
foi avaliado através do método de incorporação do extrato líquido filtrado em meio BDA. Os ensaios mostraram que
todos os MNVs produzidos inibiram de forma semelhante o fungo Sclerotinia sclerotiorum. Por outro lado, os isolados
CEN1245 e CEN1274, ambos Trichoderma brevicompactum, mostraram um amplo espectro de ação, atuando contra
Sclerotium rolfsii, Colletotrichum gloesporioides, Verticillium dahliae, Fusarium oxysporum e Cylindrocladium sp.
O presente estudo descreve isolados que produziram metabólitos não voláteis com amplo espectro de ação antifúngico,
assim como patógeno-específico. Os MNVs de Trichoderma spp. obtidos de diferentes amostras de solo cultivadas com
vegetais, mandioca e milho, foram eficientes na inibição de fungos fitopatogênicos pertencentes a outros patossistemas,
como os de espécies florestais e frutíferas, o que poderia aumentar sua potencial aplicação no controle de doenças de
plantas. Adicionalmente, estes fungos antagonistas devem ser mais bem estudados para identificação de moléculas
bioativas de interesse industrial ou formulação de produtos para o controle biológico de patógenos de plantas.
Palavras-chave: metabólitos secundários, antagonismo, inibição do crescimento micelial, fungos fitopatogênicos.

http://dx.doi.org/10.1590/1676-0611-BN-2017-0418 http://www.scielo.br/bn
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Marques, E. et al.

Introduction filtration and incorporation in the substrate method. Numerous scientific

studies have used the methodology of incorporation of filtrates into culture
Trichoderma Persoon is a hyperparasite fungus that uses different medium for the evaluation of antagonism in different plant pathogenic fungi
mechanisms of biological control, which include: parasitism, antibiosis, species, such as Verticillium dahliae Kleb. (Jamdar et al. 2013), Sclerotium
competition, induction of resistance and growth promotion (Kumar 2013). rolfsii Sacc. (Darvin et al. 2013, Isaias et al. 2014, Srinivasa et al. 2014),
Antibiosis is the mechanism by which certain microorganisms respond to the Sclerotinia sclerotiorum Lib. De Bary and Fusarium oxysporum Schlecht,
presence of others, secreting compounds or metabolites capable of inhibiting Snyder & Hansen (Jaspal et al. 2009, Castillo et al. 2011, Carvalho et al.
or preventing their development (Benítez et al. 2004). According to Dennis
2011, Saxena et al. 2014), Colletotrichum spp. (Martins et al. 2007, Ajith
& Webster (1971a, b), antibiotic compounds may be Volatile Metabolites
& Lakshmidevi 2010, Farah & Nasreen 2013) and Cylindrocladium sp.
(VMs) and Non-Volatile Metabolites (NVMs). Although these molecules are
(Carvalho Filho et al. 2008).
not essential, in some cases they are important for the selective survival of
Although this biocontrol agent has been studied for a long time,
organisms under certain conditions (Donidio & Monciardini 2002), eliminating
including in Brazil, the industry still needs new isolates with potential for
microbial competition and, consequently, leading to greater availability
use in Integrated Pest Management programs (IPM). Nowadays, according
of scarce food sources in the soil (Benítez et al. 2004). Raw metabolites
to the Brazilian Ministry of Agriculture Livestock and Food Supply
consist of mixtures of compounds resulting from primary or secondary
(MAPA), only five bioproducts, formulated with either T. asperellum and
metabolism (Yamaguchi 1996). Primary metabolites (polysaccharides,
T. harzianum, are available.
fatty acids and nucleic acids) are common to all biological systems and
Faced with the increasing demand of agriculture for ecologically
are produced in the log phase of growth. Secondary metabolites (SMs)
compatible alternatives for the management of diseases, this work aimed to
are produced during the stationary phase and in lesser quantities than the
investigate the spectrum of action of non-volatile metabolites of Trichoderma
primary ones, encompassing a diversified class of low molecular weight
isolates against different plant pathogenic fungi.
substances and produced by specific groups of organisms (Hanson 2003),
which include antibiotics.
Weidling (1941) described gliotoxin, isolated from the fungus Material and methods
Gliocladium fimbriatum Gilman & Abbott, which was one of the first
SM described and associated with antifungal action. At the present time, 1. Origin of fungal isolates
several SMs have been reported, and these are the main groups found in
Trichoderma isolates: polyketides, peptaibols, terpenoids/steroids; pyrones Eight isolates of Trichoderma from the Collection of Fungi for the
and daucanes (Zeilinger et al. 2016). The SMs may play an antibiotic Control of Plant Pathogens and Weeds of EMBRAPA (Brazilian Agricultural
function against plant pathogens (Daoubi et al. 2009, Takhiro et al. 2013), Research Corporate), already characterized and identified in previous
act as a plant growth regulator, and have pharmacological properties for studies (Marques et al. 2016), were used. According to the study cited,
use as antiaging agents, cholesterol lowering agents, cancer cell fighters, these isolates exhibited antifungal activity in dual culture tests in vitro
immunological suppressors (Keswani et al. 2014) and flavoring for the against S. sclerotiorum, and the identification was performed based on
food industry (Fadel et al. 2015). the sequencing of regions ITS1/2 of ribosomal DNA (rDNA). In addition
Levels of biological control exerted on a target organism vary according to S. sclerotiorum, six other plant pathogenic fungal isolates were used in
to the antagonistic isolate, and therefore vary both intra and interspecifically the present study, all described in Table 1.
(Dennis & Webster 1971a, b, Brodeur 2012). In the context of the
2. Evaluation of the antifungal activity
selection of these biocontrol agents, in vitro assays are used as indicative
of the mode of action of the antagonist (Carvalho Filho et al. 2008). The methodology described by Dennis & Webster (1971a) was
The evaluation of NVMs produced by fungi under laboratory conditions, used to evaluate the potential of NVMs (crude extract or filtered from
as well as dual culture, is an initial step in the prospection of metabolites cultures) produced by Trichoderma isolates against plant pathogenic fungi.
for both biological control and other purposes and can not be overlooked. Routine multiplications of both antagonists and pathogens was performed
It can be performed by the technique of cellophane paper (Khalili et al. on plastic Petri dishes containing commercial Potato-Dextrose-Agar
2012, Jeyaseelan et al. 2014, Barari 2016) or also from the liquid culture, (PDA) medium. Incubation of the cultures occurred at 25 ºC with

Table 1. Description of biocontrol agents and plant pathogenic fungi used in this study
Collection code Origin of the isolates Fungi
CEN1242 Maize (Zea mays L.) T. harzianum
CEN1245 Tomato (Solanum lycopersicum L.) T. brevicompactum
CEN1251 Maize (Z. mays) T. harzianum
CEN1255 Tomato (S. lycopersicum) T. harzianum
CEN1256 Tomato (S. lycopersicum) T. harzianum
CEN1258 Cassava (Manihot esculenta Crantz) T. spirale
CEN1270 Cassava (M. esculenta) T. spirale
CEN1274 Kale (Brassica oleracea var. acephala) T. brevicompactum
CEN216 Common bean (Phaseolus vulgaris L.) S. rolfsii
CEN419 Passion fruit (Passiflora edulis Sims) C. gloeosporioides
CEN1273 Chickpeas (Cicer arietinum L.) F. oxysporum
CEN494 Eucalyptus (Eucalyptus grandis Hill ex. Maiden) Cylindrocladium sp.
CEN788 Strawberry (Fragaria sp.) V. dahliae
CEN1147 Common bean (P. vulgaris) S. sclerotiorum

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Potential of extracts of Trichoderma

Table 2. Inhibition Index of Mycelial Growth (IMG %) of plant pathogenic fungi after being confronted with filtrates from cultures of isolates of Trichoderma
Plant pathogenic fungi
Collection code Sclerotinia Fusarium Colletotrichum Sclerotium Verticillium Cylindrocladium
sclerotiorum oxysporum gloeosporioides rolfsii dahliae sp.
CEN1242 9.7 a* 1.0 b 1.0 d 1.0 c 2.4 e 5.7 b
CEN1245 9.2 a 8.2 a 6.8 bc 9.5 a 10.2 a 5.7 b
CEN1251 8.9 a 1.0 b 1.0 d 5.0 b 2.4 e 5.0 b
CEN1270 8.9 a 1.0 b 5.1 c 5.6 b 5.5 c 5.3 b
CEN1255 9.4 a 1.0 b 1.0 d 1.0 c 1.0 f 11.0 a
CEN1256 9.5 a 1.0 b 1.0 d 1.0 c 5.2 cd 5.4 b
CEN1258 8.9 a 1.0 b 8.7 ab 1.0 c 4.3 d 5.7 b
CEN1274 9.1 a 7.7 a 9.2 a 5.7 b 9.1 b 5.9 b
CV%** 5.2 6.5 17.3 11.5 6.5 4.6
* Means followed by the same letter do not differ statistically by Tukey test (P < 0.05). ** CV: coefficient of variation.

a 12 h photoperiod for seven days. To obtain the liquid phase with the Discussion
non-volatile metabolites, the Trichoderma isolates were grown in PD
medium (Potato-Dextrose, without addition of agar) in an orbital shaker Since all isolates produced active NVMs against S. sclerotiorum, the
at 150 rpm at 25 °C with absence of light for seven days. After this period, results were compared with those obtained in previously performed dual
the liquid part was collected by filtration on filter paper and centrifuged culture tests (Marques et al. 2016), and a greater inhibition was observed
at 6.081 x g for removal of fungal spores that could hinder membrane with the use of the culture filtrates than in the direct comparison between
sterilization. The liquid phase was filtered through cellulose membranes pathogen and antagonist. This fact suggests that the main mechanism of
of 0.45 μm diameter and incorporated into the PDA medium (~ 50 °C) in action of these isolates is antibiosis, that is, the production of secondary
a proportion of 25% (v / v). Three replicates were prepared with agar discs compounds with antibiotic properties. It was observed that the three different
(5 mm diameter) taken from pathogen cultures. The mycelial agar disks species, T. harzianum, T. spirale and T. brevicompactum were effective
were deposited in the center of each Petri dish containing PDA medium, against the fungus that causes white mold (Figure 1). Saxena et al. (2014)
supplemented with the respective antagonist culture filtrates. Control reported maximum inhibition rates against this plant pathogenic fungus,
plates consisted of mycelial agar disks of each pathogen deposited in PDA using culture filtrates from T. harzianum and T. atroviride (Jaspal et al. 2009),
medium, with sterile distilled water added. while for Castillo et al. (2011) this index was higher with T. asperellum
The evaluation of the radial mycelial growth of the pathogen was NVM. It is postulated, therefore, that this action by antibiosis is variable
carried out by taking the measurements of the diameter of the colonies, intra and interspecifically, corroborating the findings of Dennis & Webster
in millimeters. These measurements were used to calculate the inhibition (1971a, b) and Brodeur (2012).
Index of Mycelial Growth (Menten et al. 1976), using the equation: The same was verified with the other plant pathogenic fungi confronted.
IMG (%) = [(Dctreat - Dtreat) / Dctreat] x100, where Dctreat = diameter of the radial Thus, the isolate that produced metabolites with the greatest capacity
mycelial growth of the pathogen in the control treatment without filtrates;
for inhibition of V. dahliea was CEN1245, belonging to the species
Dtreat = diameter of the radial mycelial growth of the pathogen in the treatment
T. brevicompactum (Table 2), while Jamdar et al. (2013) and Isaias et al.
with the filtrates. These evaluations were performed when the entire surface
(2014) observed a greater inhibitory effect against this pathogen, using filtrates
of the medium, in the control treatment, was colonized by the pathogen.
from T. harzianum, T. koningii and T. koningiopsis. With F. oxysporum,
3. Statistical analysis the NVMs that led to the greatest inhibition of mycelial growth in this
work were also those produced by T. brevicompactum (CEN1245 and
IMG data were converted to √x+1 and used in analysis of variance CEN1274). There are reports of greater inhibition of F. oxysporum
(ANOVA), followed by Tukey test, at a level of 5% probability of with metabolites of T. viride (Farah & Nasreen 2013) and T. harzianum
significance, using the program Assistat 7.6 beta (Silva & Azevedo 2016). (Farah & Nasreen 2013, Saxena et al. 2014). In addition to the filtrates from
the last two of the aforementioned isolates of T. brevicompactum, another
Results one also filtered from T. harzianum (CEN1258) inhibited the mycelial
growth of C. gloesporioides. Several studies, such as those developed by
All Trichoderma isolates tested produced NVMs that were effective
Ajith et al. (2010), Jaspal et al. (2009) and Martins et al. (2007) reported
against S. sclerotiorum, with no significant statistical difference between
the production of T. viride and T. harzianum metabolites as those with the
them (Table 2). Against F. oxysporum, only isolates CEN1245 and CEN1274
highest antifungal potential of different species of Colletotrichum, among
produced non-volatile compounds capable of inhibiting this fungus. Different
several other species tested.
inhibition indices of mycelial growth of C. gloesporioides were observed in
the treatments with NVMs produced by the isolates CEN1245, CEN1270, In the tests with Cylindrocladium sp., the isolate CEN1255, from T. harzianum,
CEN1258 and CEN1274, but without significant statistical difference between was highlighted as to the potential inhibition of growth of this pathogen.
them. The other isolates did not produce effective secondary compounds against The same was verified by Carvalho Filho et al. (2008) who, however, also
this plant pathogenic fungus. Among the isolates tested against S. rolfsii, four selected a T. pseudokoningii isolate as highly active against this pathogen
produced antifungal substances; however, it was isolate CEN1245 that stood in NVMs production tests. As for S. rolfsii, the culture filtrate with the
out. V. dahliae had mycelial growth inhibited by NVMs produced by almost highest antifungal effect was T. brevicompactum (CEN1245), while in the
all isolates tested except for CEN1255. The highest IMGs observed were, literature the filtrates were T. harzianum (Saxena et al. 2014), T. viride
once again, produced by isolates CEN1245 and CEN1274, with significant (Darvin et al. 2013) and T. virens (Srinivasa et al. 2014), corroborating,
statistical difference between them. Finally, all isolates produced SMs with once again, with Dennis & Webster (1971a, b) and Brodeur (2012).
activity against Cylindrocladium sp., although isolate CEN1255 stood out In the present study the intra and interspecific variation of the antifungal
with the highest IMG (Table 2). activity of Trichoderma isolates in relation to different plant pathogenic

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 4 Biota Neotrop., 18(1): e20170418, 2018

Marques, E. et al.

Figure 1. Effect of filtrates from cultures of Trichoderma spp. that stood out in the mycelial growth of plant pathogenic fungi, where a1-a6: Control treatments, b1-b6:
treatments with the isolate CEN1245 and c1-c5: treatments with isolate CEN1274 and a1, b1 and c1: S. rolfsii (CEN216), a2, b2 and c2: F. oxysporum (CEN1273), a3, b3
and c3: C. gloesporioides (CEN419), a4, b4 and c4: V. dahliae (CEN788), a5, b5 and c5: Cylindrocladium sp. (CEN609) and a6, b6 and c6: S. sclerotiorum (CEN1147),
respectively.

fungi was confirmed. Filtered cultures of the isolates of T. brevicompactum, References

CEN1245 and CEN1274, presented a broad antifungal spectrum, although
isolates from other species were also prominent in the production of active AJITH, P.S. & LAKSHMIDEVI, N. 2010. Effect of volatile and non-volatile
metabolites with specificity against the plant pathogenic fungi used in the compounds from Trichoderma spp. against Colletotrichum capsici incitant of
anthracnose on bell peppers. Nat. Sci. 8(9):265-269.
experiments. The Trichoderma metabolites obtained from different soil
samples cultivated with vegetables, cassava and maize were effective to BARARI, H. 2016. Biocontrol of tomato Fusarium wilt by Trichoderma species
under in vitro and in vivo conditions. Cercet. Agron. Mold. 49:91-98.
plant pathogenic fungi belonging to other patossystems such as forest or
fruit trees, which could broaden their application in the biological control BENÍTEZ, T., RINCÓN, A.M., LIMÓN, M.C. & CODÓN, A.C. 2004. Biocontrol
mechanisms of Trichoderma strains. Int. Microbiol. 7(4):249-260.
of plant diseases. In addition, the antagonistic fungi should be studied in
greater depth for the identification of bioactive molecules of industrial BRODEUR, J. 2012. Host specificity in biological control: insights from opportunistic
pathogens. Evol. Appl. 5(5):470-480.
interest or in commercial formulations of products for biological control
of plant pathogens. CARVALHO, D.D.C., MELLO, S.C.M., LOBO JUNIOR, M. & SILVA, M.C.
2011. Controle de Fusarium oxysporum f. sp. phaseoli in vitro e em sementes,
e promoção do crescimento inicial do feijoeiro comum por Trichoderma
Acknowledgements harzianum. Trop. Plant Pathol. 36(1):28-34.

The authors are grateful for the financial support of CAPES (Coordination CARVALHO FILHO, M.R., MENÊZES, J.E., MELLO, S.C.M. & SANTOS, R.P.
2008. Avaliação de isolados de Trichoderma no controle da mancha foliar do
for the Improvement of Higher Education Personnel) and FAP-DF (Federal
eucalipto in vitro e quanto a esporulação em dois substratos sólidos. Boletim
District Research Support Foundation). de Pesquisa e Desenvolvimento, Brasília, 225. 21p.
CASTILLO, F.D.H., PADILLA, A.M.B., MORALES, G.G., SILLER, M.C.,
Author Contributions HERRERA, R.R., GONZALES, C.N.A. & REYES, F.C. 2011. In vitro antagonist
action of Trichoderma strains against Sclerotinia sclerotiorum and Sclerotium
Sueli Correa Marques de Mello: substantial contribution in the cepivorum. A. J. Agric. Biol. Scie. 6(3):410-417.
concept and design of the study; contribution to manuscript preparation DAOUBI, M., PINEDO-RIVILLA, C., RUBIO, M.B., HERMOSA, R., MONTE, E.,
and critical revision. ALEU, J. & COLLADO, I.G. 2009. Hemisynthesis and absolute configuration of
Irene Martins: substantial contribution to data collection; contribution novel 6-pentyl-2H-pyran-2-one derivatives from Trichoderma spp. Tetrahedron
to data analysis and interpretation. 69:4834-40.
Eder Marques: substantial contribution in the concept and design of DARVIN, G., VENKATESH, I. & REDDY, N. 2013. Evaluation of Trichoderma
the study; contribution to data collection; contribution to data analysis and spp. against Sclerotium rolfsii in vitro. Int. J. Appl. Biol. Pharm. (4):268-272.
interpretation; contribution to manuscript preparation and critical revision. DENNIS, C. & WEBSTER, J. 1971a. Antagonistic properties of species-groups
of Trichoderma. I. Production of non-volatile antibiotics. Trans. Br. Mycol.
Conflicts of interest Soc. 57(1):25-39.
DENNIS, C. & WEBSTER, J. 1971b. Antagonistic properties of species-groups
The authors declare that they have no conflict of interest related to the of Trichoderma. II. Production of volatile antibiotics. Trans. Br. Mycol. Soc.
publication of this manuscript. 57(1):41-48.

http://www.scielo.br/bn http://dx.doi.org/10.1590/1676-0611-BN-2017-0418
Biota Neotrop., 18(1): e20170418, 2018 5

Potential of extracts of Trichoderma

DONIDIO, S. & MONCIARDINI, P. 2002. Microbial technologies for the discovery gloeosporioides. Boletim de pesquisa e desenvolvimento 193. Embrapa Recursos
of novel bioactive metabolites. J. Biotechnol. 99(3):187-198. Genéticos e Biotecnologia, Brasília, 12p.
FADEL, H.H.M., MOHMOUD, M.G. & ASKER, M.M.S. 2015. Characterization MENTEN, J.O.M., MINUSSI, C.C., CASTRO, C. & KIMATI, H. 1976. Efeito
and evaluation of coconut aroma produced by Trichoderma viride EMCC-107 in de alguns fungicidas no crescimento micelial de Macrophomina phaseolina
solid state fermentation on sugarcane bagasse. Electron. J. Biotechnol. 18(1):5-9.
(Tass.) Goid. “in vitro”. Fitopatol. Bras. 1:57-66.
FARAH, S.T. & NASREEN, S. 2013. In vitro assessment of antagonistic activity
of Trichoderma viride and Trichoderma harzianum against pathogenic fungi. MARQUES, E., MARTINS, I., CUNHA, M.O.C., LIMA, M.A., SILVA, J.B.T., SILVA,
Indian J. Appl. Res. 3(5):57-59. J.P., INGLIS, P.W., & MELLO, S.C.M. 2016. New isolates of Trichoderma
HANSON, J.R. 2003. Natural Products: The Secondary Metabolites. Royal Society antagonistic to Sclerotinia sclerotiorum. Biota Neotrop. 16(3): e20160218. http://
of Chemistry: Cambridge, pp.1772-1773. dx.doi.org/10.1590/1676-0611-BN-2016-0218 (último acesso em 17/10/2017)
ISAIAS, C.O., MARTINS, I., SILVA, J.B.T., SILVA, J.P. & MELLO, S.C.M. SAXENA, D., TEWARI, A.K. & RAI, D. 2014. In vitro antagonistic assessment of
2014. Ação antagônica e de metabólitos bioativos de Trichoderma spp. contra T. harzianum PBT 23 against plant pathogenic fungi. J. Microbiol. Biotechnol.
os patógenos Sclerotium rolfsii e Verticillium dahliae. Summa Phytopathol. Res. 4(3):59-65.
40:(1)34-41.
SILVA, F.A.S. & AZEVEDO, C.A.V. 2016. O Assistat Software Versão 7.7 e seu
JAMDAR, Z., MOHAMMADI, A.H. & MOHAMMADI, S. 2013. Study of
uso na análise de dados experimentais. Afr. J. Agric. Res. 11(39):3733-3740.
antagonistic effects of Trichoderma species on growth of Verticillium dahliae,
the causal agent of Verticillium wilt of pistachio under laboratory condition. SRINIVASA, N., DEVI, P., SUDHIRJUMAR, S., KAMIL, D., BORAH, J.L. &
J. Nuts 4(4):53-56. PRABHAKARAN. 2014. Bioefficacy of Trichoderma isolates against soil-
JASPAL, K., MUNSHI, G.D., SINGH, R.S. & KOCK, E. 2009. Selection of borne pathogens. Afr. J. Microbiol. Res. 8(28):2720-2723.
biocontrol agents for the management of white rot of peas caused by Sclerotinia WEIDLING, R. 1941. Experimental consideration of the mold toxins of Gliocladium
sclerotiorum. Plant. Dis. Res. 24(2):148-155. and Trichoderma. Phytopathol. 27:991.
JEYASEELAN, E.C., THARMILA, S. & NIRANJAN, K. 2014. Antagonistic activity
ZEILINGER, S.Z., GRUBER S., BANSAL R. & MUKHERJEE P. 2016. Secondary
of Trichoderma spp. and Bacillus spp. against Pythium aphanidermatum isolated
metabolism in Trichoderma – Chemistry meets genomics. Fungal Biol. Rev.
from tomato damping off. Arch. Appl. Sci. Res. 4(4):1623-1627
30(2):74-90.
KHALILI, E., SADRAVI, M., NAEIMI, S. & KHOSRAVI, V. 2012. Biological
control of rice brown spot with native isolates of three Trichoderma. Braz. J. YAMAGUCHI, I. 1996. Pesticides of microbial origin and application of molecular
Microbiol. 43(1):297-305. biology. In: Copping, L.G. (eds.), Crop Protection Agents from Nature: Natural
KESWANI, C., MISHRA, S., SARMA, B.K., SINGH, S.P. & SINGH, H.B. 2014. Products and Analogues. Royal Society of Chemistry, Cambridge, UK. pp.27-49.
Unraveling the efficient applications of secondary metabolites of various
Trichoderma spp. Appl. Microbiol. Biotechnol. 98:533-544.
Received: 02/08/2017
KUMAR, S. 2013. Trichoderma: A biological weapon for managing plant diseases
and promoting sustainability. Int. J. Agrl. Sc. & Vet. Med. 1(3):1-18. Revised: 14/12/2017
MARTINS, I., PEIXOTO, J.R., MENÊZES, J.E. & MELLO, S.C.M. 2007. Accepted: 15/12/2017
Avaliação in vitro do antagonismo de Trichoderma spp. sobre Colletotrichum Published online: 01/02/2018

http://dx.doi.org/10.1590/1676-0611-BN-2017-0418 http://www.scielo.br/bn