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Appl Microbiol Biotechnol (2014) 98:533–544

DOI 10.1007/s00253-013-5344-5

MINI-REVIEW

Unraveling the efficient applications of secondary metabolites


of various Trichoderma spp.
Chetan Keswani & Sandhya Mishra &
Birinchi Kumar Sarma & Surya Pratap Singh &
Harikesh Bahadur Singh

Received: 15 July 2013 / Revised: 16 October 2013 / Accepted: 17 October 2013 / Published online: 27 November 2013
# Springer-Verlag Berlin Heidelberg 2013

Abstract Recent shift in trends of agricultural practices from generate greater understanding of their other important and
application of synthetic fertilizers and pesticides to organic beneficial applications apart from target specific biopesticides.
farming has brought into focus the use of microorganisms that
carryout analogous function. Trichoderma spp. is one of the Keywords Trichoderma . Secondary metabolites . Biological
most popular genera of fungi commercially available as a control . Antibiotics
plant growth promoting fungus (PGPF) and biological control
agent. Exploitation of the diverse nature of secondary metab-
olites produced by different species of Trichoderma augments Introduction
their extensive utility in agriculture and related industries. As a
result, Trichoderma has achieved significant success as a Over the past three decades, fungicolous fungi have attracted
powerful biocontrol agent at global level. The endorsement global interest for their biocontrol potential against phytopath-
of Trichoderma spp. by scientific community is based on the ogens especially in organic farming. Globally, Trichoderma
understanding of its mechanisms of action against a large set occupies the top slot in the segment of fungal biocontrol
of fungal, bacterial and in certain cases viral infections. agents (Whipps and Lumsden 2001). The biocontrol potential
However, it is still an agnostic view that there could be any of Trichoderma has been the most fascinating and interesting
single major mode of operation, although it is argued that all topic of research for agricultural scientists. But since the initial
mechanisms operate simultaneously in a synchronized fash- investigation by Weindling (1932) regarding the antagonistic
ion. The central idea behind this review article is to emphasize nature of Trichoderma spp., a remarkable discovery has been
the potentiality of applications of target specific secondary made of late after the genome profiling of three commercially
metabolites of Trichoderma for controlling phytopathogens important species of Trichoderma: T. virens, T. atroviride and
as a substitute of commercially available whole organism T. reesei (Mukherjee et al. 2012). This has opened new
formulations. With the aim to this point, we have compiled avenues for understanding the molecular basis of biological
an inclusive list of secondary metabolites produced by differ- control mechanisms of Trichoderma (Kubicek et al. 2011).
ent species of Trichoderma and their applications in diverse The global scientific community has shown keen interest in
areas with the major emphasis on agriculture. Outlining the Trichoderma spp. owing to its broad-spectrum antagonistic
importance and diverse activities of secondary metabolites of activities against various soil borne phytopathogens (Singh
Trichoderma besides its relevance to agriculture would et al. 2011). The major modes of action include: (a)
mycoparasitism — direct confrontation with mycopathogens,
secretion of cell wall degrading enzymes, subsequent penetra-
C. Keswani : S. Mishra : B. K. Sarma : H. B. Singh (*)
tion and killing are the key features of this mode (Woo and
Department of Mycology and Plant Pathology, Institute of
Agricultural Sciences, Banaras Hindu University, Varanasi 221005, Lorito 2007); (b) antibiosis — secretion of secondary metab-
India olites having antimicrobial properties, in the vicinity of its
e-mail: hbs1@rediffmail.com niche (Sivasithamparam and Ghisalberti 1998); (c) competi-
tion — being decomposers by nature, Trichoderma spp. effi-
C. Keswani : S. P. Singh
Department of Biochemistry, Banaras Hindu University, ciently mobilize and uptake macro- and micro-nutrients from
Varanasi 221005, India soil, which results in scarcity of nutrients for other microbes in
534 Appl Microbiol Biotechnol (2014) 98:533–544

its vicinity leading to decreased inter and intra species com- Table 1 List of Trichoderma secondary metabolites exhibiting multiple
applications
petition (Benitez et al. 2004; Verma et al. 2007).
Though it is difficult to suggest which is the major mode of Compounds Multiple applications References
operation, Di Pietro et al. (1993) and Schirmbock et al. (1994)
have suggested that Trichoderma spp. operates mainly through Emodin Cathartic stimulant, tumor Ali et al. 2004; Huang
cell adhesion inhibitor et al. 2006
synchronization between mycoparasitism and antibiosis
Gliotoxin Antimalarial, immune Pahl et al. 1996; Tanaka
against the fungal pathogens. It would be important to note system suppressor et al. 1998
that Trichoderma spp. constitutively secrete a variety of lytic
Harzianolide Antifungal, plant growth Vinale et al. 2006, 2008
enzymes to detect the presence of a competent fungi by sensing promoter
their degraded cell wall components (Harman et al. 2004; Woo Koninginins Antifungal, plant growth Cutler et al. 1991a;
and Lorito 2007; Vinale et al. 2008), while no such reports for regulator Ghisalberti and
Rowland 1993, Parker
constitutive secretion of antimicrobial secondary metabolites
et al. 1995a, b
in absence of a potential competitor are available. Thus, it
6-Pentyl-2H- Antifungal (plant pathogen), Claydon et al. 1987;
would be difficult to comment on the role of secondary me- pyran-2-one plant growth promoter, Parker et al. 1997;
tabolites in sensing the pathogens but as soon as the pathogen coconut aroma Vinale et al. 2008
has been sensed, secretion of antimicrobial secondary metab- Trichokonins Broad spectrum antifungal, Xiao-Yan et al. 2006;
olites is triggered (Vinale et al. 2009b). It is a well known fact plant defense inducer Luo et al. 2010
that Trichoderma spp. secretes a variety of secondary metab- Viridiofungins Potential anticancer Harris et al. 1993;
compound, bactericidal Mandala et al. 1997
olites that not only contribute significantly in signaling but also
interact with other organisms in various ways. Although re- Viridin Broad spectrum antifungal Brian and McGowan
compound, anti- 1945; Dodge et al.
searchers mostly focused their attention on the immense po- neoplastic, anti- 1995
tentiality of Trichoderma secondary metabolites in biological atherosclerosis
control of phytopathogens, the fact that other secondary me- Viridiol Weedicidal, anti-aging Sakuno et al. 2000;
tabolites also exhibit multiple biological functions cannot be Heraux et al. 2005b
ignored. Therefore, a representative list of secondary metabo-
lites with potential multifarious biological activities such as
plant growth regulation, weedicidal effect, antibiotic, anti- a broad range of phytopathogens. Undoubtedly, the chemical-
aging, antiplasmodial, cholesterol lowering agent, flavoring ly diverse type of secondary metabolites produced by
agent, and immune suppressor is provided in Table 1. Trichoderma is the source of its efficient biocontrol potential.
Prior art is loaded with information concerning various Multiple strategies have been adapted for development of
secondary metabolites produced by Trichoderma spp. biopesticide formulation of Trichoderma viz. whole organ-
However, limited information is available on the molecular ism, spore suspension and talc-based formulations, which
mechanisms of these secondary metabolites in inter- and intra- proved successful in controlling various plant pathogens.
species interactions. Here, we tried to appraise the variability However, such kinds of formulations do exhibit certain draw-
among different Trichoderma spp. in fabricating a wide array backs which need to be addressed for making Trichoderma-
of secondary metabolites. We also attempted to update the based biopesticide a luminary in pesticide enterprises. These
contemporary facts based on species wise classification of issues include limited shelf life, high dose requirement, effec-
secondary metabolites secreted by various non-pathogenic tiveness against soil/seed borne pathogens only, and low
and industrially valuable Trichoderma spp. and have also stability in adverse environment. For instance, Trichoderma
logically pointed out their possible prospecting for commer- spp. has a narrow application range for majority of phytopath-
cial applications in agriculture, pharmacy and food industries ogenic bacteria in comparison to fungal pathogens because
(Fig. 1, Table 2). In addition, one of our main interests is to bacteria generally have relatively higher metabolic rates than
improve or replace the conventional formulations of biopesti- fungi, thus reducing its effectiveness against bacterial patho-
cides so that next-generation secondary metabolites based gens (Verma et al. 2007). However, Fravel (1988) reported
formulations could be developed for management of that purified secondary metabolites from Trichoderma spp.
phytopathogens. could be potentially effective in controlling bacterial infec-
tions more rapidly than whole organism application under
field conditions. Furthermore, it is interesting to note that
Trichoderma secondary metabolites: impact on biological while using Trichoderma spore formulation, two mechanisms
control of phytopathogens operate parallel to each other firstly, simultaneous secretion of
cell wall degrading enzymes and antimicrobial secondary
Trichoderma is lucratively used as potent biocontrol agent metabolites which ensures pathogen killing by either way
worldwide due to its prominent antimicrobial activity against and secondly, a simultaneous increase in Trichoderma
Appl Microbiol Biotechnol (2014) 98:533–544 535

Fig. 1 Multi-farious role of


Trichocaranes, Koninginins ,
Trichoderma secondary
Viridiol, (3H)-benzoxazolinone , Trichosetin, Cyclonerodiol ,
metabolites
2,4-dihydroxy -1,4-(2H)- Harzianolide, Trichokonin ,
benzoxazine-3-one Viridiol, 6-pentyl-2H-pyran-2-
one
Weedicidal Plant Growth
Regulators

6-pentyl-2H-pyran-2-
one, 3-octanone, 1- R- mevalonolactone,
Flavour and Anti-aginig
Aroma Mevastatin
octen-3-ol, 3-octanol

Anti - cancer Biological Control


Viridepyrone,
Trichodenones, Massoilactone, Koninginins ,
Trichodermamide B, Viridin, Viridiofungins,
Harzianum A, Aurocitrin, Harzianopyridone ,
Harziphilone, Viridin, Virone, T22- azaphilone, Harzianolide,
Viridiofungin Trichostromaticins, Harzianic
acid

population in the field which is a major agenda of sustainable 1991; Vinale et al. 2006). The effectiveness of the same
agriculture (Fig. 2). metabolites at concentrations between 1 and 10 mg/ml in
Trichoderma spp. secretes a chemically diverse range of reducing the incidence of B. cinerea and Leptosphaeria
secondary metabolites, of which broad-spectrum antimicrobi- maculans on tomato and canola seedlings was further proved
al properties have been demonstrated well in many in vitro in planta (Vinale et al. 2008). Furthermore, Vinale et al. (2011)
assays. The volatile compound 6-pentyl-2H -pyran-2-one is showed 100 %, 41 % and 28 % inhibition of P. ultimum , R.
active against Rhizoctonia solani and Fusarium oxysporum f. solani and B. cinerea, respectively, at 100 μg/plug concen-
sp. lycopersici (Scarselletti and Faull 1994). Moreover, in tration of cerinolactone. Andrade et al. (1992) reported the
another study its application was also found to reduce rotting antagonistic effects of 5-hydoxyvertinolide against Mycena
of stored kiwi fruits caused by Botrytis cinerea (Poole et al. citricolor which causes American leaf spot of coffee, whereas
1998). Likewise, viridepyrone showed antagonistic activity koningic acid has specific activity against anaerobic bacteria
against Sclerotium rolfsii , whereas massoilactone and Bacteroides fragilis (Itoh et al. 1980). TAEMC161 re-
δ-decanolactone were found to control B. cinerea, identified as viridiol is a potent enzyme inhibitor, inhibiting
Phytophthora spp., Aspergillus niger and Candida albicans the activity of 5′-hydroxyaverantin dehydrogenase which cat-
(Hill et al. 1995; Evidente et al. 2003; Kishimoto et al. 2005). alyzes an early step in aflatoxin biosynthesis in Aspergillus
Complex pyrones such as koninginins A and B have shown flavus and A. parasiticus (Sakuno et al. 2000; Wipf and
antifungal activity against take all fungus Gaeumannomyces Kerekes 2003). Crown and stem rot of artichoke caused by
graminis var. tritici (Ghisalberti and Rowland 1993), while S. rolfsii was considerably inhibited by viridepyronone at
koninginin D inhibited the growth of Bipolaris sorokiniana, concentration 196 μg/ml (Evidente et al. 2003).
Pythium middleonii, F. oxysporum, Phytophthora cinnamomi Trichostromaticins A–E in field conditions have shown sig-
and R. solani (Dunlop et al. 1989). Similarly, viridin, a broad nificant results in controlling Moniliophthora perniciosa ,
spectrum antifungal compound, prevented spore germination which causes cocoa witches broom disease in South
of Stachybotrys atra , Aspergillus niger , Penicillium America (Degenkolb et al. 2008). Another compound
expansum , Colletotrichum lini , Fusarium caeruleum , and harzianic acid at 10 μg showed complete inhibition of
Botrytis allii (Brian and McGowan 1945), whereas Pythium irregulare and S. sclerotiorum while at 100 μg
viridiofungins have shown microbicidal activity against completely arrested the growth of R. solani (Vinale et al.
Candida, Aspergillus, and Cryptococcus spp. (Harris et al. 2009a). Meanwhile, Staphylococcus aureus, Bacillus subtilis,
1993). Application of racemic form of harzianopyridone Streptococcus faecalis , Clavibacter michiganensis ssp.
showed potent antifungal activity against R. solani, B. cinerea michiganensis, F. oxysporum f. sp. phaseoli, F. oxysporum
(Dickinson et al. 1989), Pythium ultimum and G. graminis f. sp. niveum, F. oxysporum f. sp. vasinfectum , B. cinerea, R.
var. tritici (Vinale et al. 2006). Secondary metabolites such as solani , Curvularia lunata , Bipolaris sorokiniana and
T22-azaphilone, harzianolide and T39 butenolide from spe- Colletotrichum lagenarium have all been reported to be sen-
cific Trichoderma strains have shown in vitro inhibition of R. sitive to trichokonins VI, VII and VIII in agar disk diffusion
solani, P. ultimum and G . graminis var. tritici (Almassi et al. assay (Xiao-Yan et al. 2006).
536 Appl Microbiol Biotechnol (2014) 98:533–544

Table 2 Species-wise classification of secreted secondary metabolites of Trichoderma

Speciesa Secondary metabolites Reference

T. aggresivum Mellein Krupke et al. (2003)


T. album 5-Hydroxy-3-methoxy-5-vinylcyclopent-2-en-1-one, Strunz et al. (1977), Sivasithamparam and Ghisalberti (1998)
Pencolide, 3-methoxy-5-hydroxy-5-allyl cyclopentenone
T. atroviride Atroviridins A, B and C, Ferricrocin, α-phellandrene, Oh et al. (2000), Vinale et al. (2008), Stoppacher et al. (2010)
α-terpinene, β-phellandrene, β-bisabolene,
α-bergamontene, α-curcumene, γ- curcumene
T. aureoviride Pachybasin, Chrysophanol, Koninginin G De Stefano and Nicoletti (1999), Cutler et al. (1999)
T. cerinum Harzianolide, Dehydroharzianolide, T39butenolide, Vinale et al. (2011)
Cerinolactone
T. citrinoviride Trichotetronine, 16,17-dihydrobislongiquinolide, Balde et al. (2010)
Dihydrotrichodimerol
T. hamatum Viridiol, Methyl ester derivative of Dermadin, Isonitric Hussain et al. (1975), Brewer et al. (1979), Fujiwara et al.
acid E and F, Isonitrins A, B, C and D, Spirolactone, (1982), Baldwin et al. (1985), Boyd et al. (1991),
Mannitol, Gliotoxin, Ergosterol, TAEMC 161 Sivasithamparam and Ghisalberti (1998), Sakuno et al.
{4,6-dihydroxy-5-methoxy-6a-methylcyclohexa[de] (2000)
indano-[7,6-e]cyclopenta[c]2H-pyran-1,9-dione},
Epoxydiol, Diol isocyanide, Tetrhydroxy isocyanide
T. harzianum Harzianic acid, Dimethyl-harzianic acid, Homoharzianic Fujiwara et al. (1982), Baldwin et al. (1985), El Hajji et al. (1987),
acid, 1-hydroxy-3-methyl-anthaquinone, 1,8-dihydroxy- Dickinson et al. (1989), Rebuffat et al. (1989), Almassi et al.
3-methyl-anthraquinone, Harzianolide, Trichoharzin, (1991), Claydon et al. (1991), Ghisalberti et al. (1992),
Harziandione, 6-pentyl-2H-pyran-2-one, 6-(1′-pentenyl)- Kobayashi et al. (1993), Ghisalberti and Rowland (1993),
2-H-pyran-2-one, Koninginin A, B and E, Cyclonerodiol, Sawa et al. (1994), Iida et al. (1994), Lee et al. (1995a,b),
Harzianopyridone, Melanoxadin, Trichodenones A, B Iida et al. (1995), Rebuffat et al. (1995), Huang et al.
and C, Harziphilone, Fleephilone, T22azaphilone, (1995a, b), Qian-Cutrone et al. (1996), Lee et al. (1997),
Harzialactones A and B, R-mevalonolactone, Harzianolide, Tezuka et al. (1997), Parker et al. (1997), Amagata et al.
T39butenolide, MR304A, MR566B, Trichosetin, (1998), Marfori et al. (2002), Kawada et al. (2004), Combet
Trichorzianines A, B, Trichokindins I- VII, Harzianins et al. (2006), Vinale et al. (2006), Vinale et al. (2009a, b)
HC, Trichorozins I–IV, 15-hydroxyacorenone,
Isonitrin A and D.
UV-induced Homothallin II Faull et al. (1994)
mutant
T. koningii 6-Pentyl-2H-pyran-2-one, Koninginin A, B, C, D and E, Tamura et al. (1975), Nobuhara et al. (1976), Itoh et al. (1980),
Viridin, Dermadin, Isonitrile trichoviridin, Homothallin I Simon et al. (1988), Edenborough and Herbert (1988), Cutler
and II, Ergokonin A and B, Cyclonerodiol, Trichokonins et al. (1989), Dunlop et al.(1989), Reichenbach et al. (1990),
V–VIII, Trikoningin KA and KB, Koningic acid, Cutler and Jacyno (1991), Augustiniak et al. (1991), Cutler
Trichoacorenol, Cyclo (L -Pro-L -Leu), methoxybenzoate, et al. (1991a,b), Auvin-Guette et al. (1993), Parker et al.
Coccinol, Tri-acorenol, Ceramide, p-hydroxybenzyl alcohol (1995a,b), Huang et al. (1995a, b), Mukhopadhyay et al.
(1996)
T. lignorum Lignoren Berg et al. (2004)
T. longibrachiatum 5-Hydroxyvertinolide, Epoxysorbicillinol, Trichodimerol, Endo et al. (1985), Nakano et al. (1990), Rebuffat et al. (1991),
Bisvertinolone, Bisvertinol, Sorbicillin, Trichodermolide, Andrade et al. (1992), Auvin-Guette et al. (1992, 1993), Iida
Sorbiquinol, Bislongiquinolide, Trichodermolide, et al. (1993), Andrade et al. (1996, 1997), Sperry et al. (1998),
Sorbiquinol, Analogues of Gliovirin {DC1149B, Vicente et al. (2001), Leclerc et al. (2001), Mohamed-Benkada
DC1149R}, Ergokonin A, Trichogins A, Polysporins et al. (2006), Degenkolb et al. (2006)
A, B, C and D, Trichosporin B-V, Trichobrachin A I–IV;
B I–IV, Tricholongins B I, II and III, Longibrachin
LGB II and III, Trichogin GA IV, Mevastatin
T. polysporum Chrysophanol, Emodin, Pachybasin, Trichopolyns I and II, Fujita et al. (1981), (1984), Donnelly and Sheridan (1986),
Cyclonerodiol oxide, Epicyclonerodiol oxide, Valinotricin, Mihara et al. (1994), Degenkolb et al. (2006)
Egosterol, Trichosporin B-IVc
T. pseudokoningii Lanosterol, Ergosterol, Pyrocalciferol, Mevastatin, 2′,4′-di- Kamal et al. (1971), Endo et al. (1985), Watts et al. (1988),
hydroxy-3′-methoxymethyl-5′-methylacetophenone, Sivasithamparam and Ghisalberti (1998), Astudillo et al.
Clavatol, Trichokonin, 2,5-dimethylbenzoquinone, (2000), Luo et al. (2010)
Epifriedelinol, Itaconic acid, Trichodermene A, Methyl-2,
4, 6-octatriene carboxylate, 2-hydroxymalonic acid
T. reesei Paracelsin, Ferricrocin Brueckner et al. (1984), Watts et al. (1988), Kubicek et al. (2011)
T. saturnisporum Paracelsin A, B, C, D and E Rebuffat et al. (1993), Ritieni et al. (1995)
T. stromaticum Trichostromaticins A, B, C, D and E Degenkolb et al. (2006)
T. strigosum Lipostrigocin A1–A6, B1–B9, Tricholongin B I, II and III Degenkolb et al. (2006)
Appl Microbiol Biotechnol (2014) 98:533–544 537

Table 2 (continued)

Speciesa Secondary metabolites Reference

T. virens L -735,334
{carotene type metabolite}, Trichocaranes A, B, C Brian and McGowan (1945), Stipanovic and Howell (1982),
and D, Trichodermamide A and B, Viridin, Wortmannolone, Blight and Grove (1986), Jalal et al. (1987), Lee et al.
Gliovirin, Ferricrocin, cis-fusarinine, trans-fusarine, (1995a,b), Macias et al. (2000), Garo et al. (2003),
Dimerum acid, Coprogen B Singh et al. (2005), Kubicek et al. (2011)
T. viride Pachybasin, Chrysophanol, Emodin, 6-pentyl-2H-pyran- Brian (1944), Wright (1954), Godtfredsen and Vangedal
Brown conidiating 2-one, Viridepyronone, Viridin, Viridiol, Viridiofungins (1964), Pyke and Dietz (1966), Slater et al. (1967), Moffatt
mutant (UV) A, B, C, A1–4, B2 and Z2, Dermadin, Isonitrile trichoviridin, et al. (1969), Yamano et al. (1970), Collins and Halim
Gliotoxin, Ergokonin A, Suzukacillin A, Trichovirins II, (1972), Golder and Watson (1980), Itoh et al. (1980), Harris
Trichorovins Ia, Ib, a, IIb, IIc, IIIa, IVa, IVb, V and VIb, et al. (1993), Kumeda et al. (1994), Fujita et al. (1994),
Trichodecenins I and II, Trichocellins, Koningic acid, Mandala et al. (1997), Sivasithamparam and Ghisalberti
1-hydroxy-3-methylanthraquinone, 1,8-dihydroxy-3- (1998), Jaworski et al. (1999), Evidente et al. (2003),
methylanthraquinone, 1,6,8-trihydroxy- Krause et al. (2006), Degenkolb et al. (2006)
3methylanthraquinone Ghisalberti (2002)
1,3,6,8-Tetrahydroxyanthraquinone, 1-acetyl-2,4,5,
7-tetrahydroxyanthraquinone
a
The historic species concept established by Rifai (1969) are currently replaced by phylogenetic concepts based on rDNA/ITS sequencing (Kullnig et al.
2001; Hermosa et al. 2004; Druzhinina et al. 2006)

Impact on plant growth regulation l showed more vigorous growth and an extensive root
system (Vinale et al. 2008).
A plethora of chemically diverse secondary metabolites secret- Treatment with harzianolide at a concentration of 1 mg/l
ed by Trichoderma spp. have been reported as both plant has positive effect on the growth and defense response in
growth promoters as well as inhibitors (Vinale et al. 2008, Brassica napus and Solanum lycopersicum seedlings
2009b; Luo et al. 2010). Trichocaranes A and B have shown (Vinale et al. 2008). In canola seedlings inhibition up to
inhibitory effect in etiolated wheat coleoptiles up to 40 % at 45 % and 33 % in stem length was reported on application
10−4 M concentration, while trichocarane C inhibited growth of harzianic acid at concentration of 100 and 10 μg/seed,
up to 86 % at 10−3 M concentration (Macias et al. 2000). respectively, while at concentration of 100, 10, 1 ng/seed,
Similarly, koninginins B and C have also shown complete stem length increased by 42 %, 44 %, 52 %, respectively, in
growth inhibition of etiolated wheat coleoptiles at 10−3 M comparison to control (Vinale et al. 2009a). Trichokonin has
concentration (Cutler et al. 1991a; Parker et al. 1995a) while proven its activity in inducing defense responses in Nicotiana
koninginins E and G showed 65 % and 56 % growth inhibition tabacum against tobacco mosaic virus (TMV) infection.
of etiolated wheat coleoptiles, respectively, at 10−3 M concen- Foliar treatment of trichokonin at concentration of 100 nM
tration (Cutler et al. 1989; Parker et al. 1995b). Marfori et al. led to 54 % lesion inhibition, 57 % reduction in average lesion
(2002) reported trichosetin (10 μg/ml) as an inhibitor of root diameter and 30 % reduction in average lesion area in system-
and shoot growth in seedling growth assays of Oryza sativa ic tissue (Luo et al. 2010).
(87 % and 32 %), Vigna radiata (67 % and 68 %, respective- In an interesting study, herbicidal activity of Trichoderma
ly), Medicago sativa (82 % and 81 %, respectively), Capsicum virens composted chicken manure and rye cover crop was also
frutescens (93 % and 67 %, respectively), and Lycopersicum observed against Setaria viridis and Amaranthus retroflexus
esculentum (91 % and 88 %, respectively). (broad leaf and grass weeds). The active compounds demon-
The laevorotatory form of harzianopyridone displayed strating herbicidal activity were recognized as viridiols, (3H)-
significant cytotoxicity in etiolated wheat coleoptiles bio- benzoxazolinone and 2,4-dihydroxy-1,4-(2H)benzoxazine-3-
assay and caused complete inhibition at 10−3 M concentra- one released during composting of manure (Heraux et al.
tion. It also caused necrosis in bean, tobacco and corn in a 2005a; Javaid and Ali 2011). Interestingly, T. virens was dem-
concentration dependent manner (Cutler and Jacyno 1991). onstrated to contribute actively in the composting process
Similarly, cyclonerodiol inhibited the growth of wheat co- through secretion of these secondary metabolites. Additionally,
leoptiles up to 61 % at 0.001 M concentration (Cutler et al. Javaid and Ali (2011) also illustrated herbicidal potential of four
1991b). Interestingly, seedling growth assay of 6-pentyl- Trichoderma species viz. T. harzianum Rifai, T. pseudokoningii
2H -pyran-2-one demonstrated inhibitory effects at high Rifai, T. reesei Simmons and T. viride Pers. against Rumex
concentration and growth promotion at low concentration dentatus, an important and noxious weed of wheat by foliar
in wheat seedlings. However, in foliar spray treatment on application. These four Trichoderma species significantly re-
tomato, the same compound at the concentration 0.166 mg/ duced the root and shoot biomass of the same weed.
538 Appl Microbiol Biotechnol (2014) 98:533–544

Fig. 2 Trichoderma antibiosis for control of soil borne phytopathogens

Impact on human health viridiofungin inhibits the enzyme farnesyl transferase which
catalyzes the farnesylation of oncogenic Ras protein pointing
An array of secondary metabolites from Trichoderma spp. has their potential in cancer treatment (Meinz et al. 1993). Viridin,
shown prominent activity against many modern age human wortmannolone and virone are inhibitors of the enzyme
diseases and their commercialization could make mankind a inositol-3-kinase which plays a major role in cell signaling
healthier species. Trichodenone A, B and C have shown pathways and can be used for treatment of neoplasm, resteno-
significant cytotoxicity against P388 lymphocytic leukemia sis and atherosclerosis (Dodge et al. 1995).
cell lines with ED50 (median effective dose) of 0.2, 1.21 and Trichodimerol demonstrated significant inhibitory activity
1.45 μg/ml, respectively (Amagata et al. 1998). In addition, against lipopolysaccharide induced production of tumor ne-
trichodermamide B was observed to be cytotoxic against crosis factor-α (TNF-α) in human monocytes and could be
HCT-116 human colon carcinoma with IC50 (half-maximal very useful in treatment of septic shock (Mazzucco and Warr
inhibitory concentration) of 0.32 μg/ml under in vitro condi- 1996). Similarly, dihydrotrichodimerol and trichotetronine al-
tion. While harzianum A has shown cytotoxicity against so suppress the production of TNF-α and nitric oxide in
HT1080 and HeLa cell lines with IC50 values of 0.65 and lipopolysaccharide stimulated RAW264.7 cells. These com-
5.07 μg/ml, respectively (Lee et al. 2005), aurocitrin exhibited pounds also activate peroxisome proliferator activated
moderate antiplasmodial activity and cytotoxicity against receptor-Y (PPAR-Y) which plays a key role in cancer cell
three cancer cell lines, KB (oral human epidermoid carcino- biology (Lee et al. 2005). Moreover, these compounds exhib-
ma), BC (human breast cancer), and NCI-H187 (human small ited growth inhibitory activity (IC50 <100 μM) in five human
cell lung cancer) having IC50 value of 4.2, 2.5 and 0.87 μM, cancer cell lines namely U373 glioblastoma, A549 non-small
respectively (Berkaew et al. 2008). Similarly, harziphilone cell lung cancer, SKMEl-28 melanoma cancer, OE21 esoph-
was also demonstrated to have in vitro cytotoxicity against ageal cancer, Hs683 oligodendroglioma and one mouse can-
murine tumor cell line M-109 at 38 μM concentration, while cer cell line B16F10 melanoma (Balde et al. 2010).
another analogous compound fleephilone inhibits the binding Emoidin with antineoplastic activity (Huang et al. 2006)
of the virion expression protein (REV) to REV-responsive and bacteriostatic effect towards S. aureus at 10 μg/ml (Wu
element in HIV infection with IC50 value of 7.6 μM (Qian- et al. 2006) also acts as a cathartic agent (Ali et al. 2004).
Cutrone et al. 1996). While lignoren when used in common agar well diffusion
In an interesting study, Yamashita (2000) showed the anti- assay at concentration of 100 μg/well showed moderate anti-
aging effects of R-mevalonolactone by activation of choles- microbial activity towards B. subtilis , Mycobacterium
terol metabolism in skin of aged humans. Another compound smegmatis , Pseudomonas aeruginosa , Sporoblomyces
compactin renamed as mevastatin (Endo et al. 1985) also acts salmonicolor and Rhodotorula rubra (Berg et al. 2004). A
as a cholesterol lowering agent by inhibiting the rate limiting broad spectrum antifungal compound chrysophanol, was
step of cholesterol biosynthesis catalyzed by enzyme HMG found to be active against C. albicans (50 μg/ml),
CoA reductase (Goldstein et al. 1979). Furthermore, Cryptococus neoformans (50 μg/ml), Trichophyton
Appl Microbiol Biotechnol (2014) 98:533–544 539

mentagrophytes (25 μg/ml) and Aspergillus fumigatus The Trichoderma toxin phobia
(50 μg/ml) (Agarwal et al. 2000). A carotane type metabolite
CAF-603 has shown antifungal activity against different There are increasing reports which present Trichoderma as an
strains of C. albicans (0.8–12.5 μg/ml), Torulopsis glabrata agriculturally imperative microorganism with extensive in-
(50 μg/ml) and A. fumigatus (25 μg/ml) (Watnanabe et al. dustrial applications. But it is interesting to note that the genus
1990). Trichoderma also comprises species performing as opportu-
Pahl et al. (1996) reported gliotoxin mediated inhibition of nistic human pathogens. The pathogenic behaviors of these
transcription factor NF-κB, a central mediator in cell mediated species are determined by particular kind of secondary me-
immune response. In addition to this, Tanaka et al. (1998) tabolites produced by brevicompactum clade (Nielsen et al.
demonstrated anti-malarial properties of gliotoxin (0.01 μg/ 2005). Noteworthy, the brevicompactum clade is recognized
ml) against merozoites and schizonts. Another compound as a separate lineage in genus Trichoderma, which includes T.
trichodermol has also shown antiplasmodial activity arundinaceum, T. turrialbense, T. protrudens and Hypocrea
(Takashima and Wataya 1999). While, a formulation of sec- rodmanii (Degenkolb et al. 2008). All the listed species of this
ondary metabolites of Trichoderma viride VCRC F24 has clade are reported to produce trichotoxins like trichodermin
shown remarkable oviposition response against female and harzianum A, B (classified as trichothecene type toxins)
Culex quinquefasciatus mosquito, which is the vector of (Nielsen et al. 2005; Degenkolb et al. 2008). Likewise,
lymphatic filariasis caused by the nematode Wuchereria Trichoderma spp. produces peptide antibiotics rich in Aib
bancrofti. Hence, this kind of formulation can be very useful (α-aminoisobutyric acid) which are classified as peptaibols.
in vector management (Geetha et al. 2003). In particularly, in vitro assays suggest that peptaibols like
alamethicins, paracelsin and trichobrachins are highly toxic
to invertebrate models Crassostrea gigas, Artemia salina and
Applications in food, flavor and aroma industry Daphnia magna (Favilla et al. 2006; Poiriera et al. 2007).
Degenkolb et al. (2008) through an extensive phylogenetic
Besides functional application in plant growth promotion, analysis have shown that brevicompactum clade is not closely
plant disease control and human curative effects, related to the species that have biological applications. Thus,
Trichoderma secondary metabolites were also found to have only members of species other than those included in
extensive application in food, flavor and aroma industries. Our brevicompactum clade could be efficiently used in biological
earlier reports have explored the possible perspective of control of soil-borne mycopathogens in field conditions.
Trichoderma in maintaining biosecurity owing to its bioactive
metabolites and tendency to modulate antioxidants potential
(Singh and Singh 2009; Singh et al. 2010). With the growing
Conclusion
interest in this area, the precise role of Trichoderma secondary
metabolites has been elaborated. The presence of volatile and
The reputation of Trichoderma spp. in the scientific commu-
non-volatile compounds in Trichoderma influences its utility
nity and industry is based on its twin advantages of plant
in enhancing mainly flavor and aroma of the food products. A
volatile compound 6-pentyl-2H -pyran-2-one imparts coconut
aroma in viable Trichoderma cultures and is significantly
used as a coconut flavoring agent in confectionary (Claydon
et al. 1987). The solid state fermentation using bagasse and
desiccated coconut is the most accepted technology for en-
hanced 6-pentyl-2H -pyran-2-one production (Sarhy-Bagnon
et al. 2000). Likewise, 3-octanone, 1-octen-3-ol and 3-octanol
produced by Trichoderma spp., contribute to mushroom fla-
vor and aroma (Combet et al. 2006). Earlier Janssens et al.
(1992) reported the methyl ketone producing ability of T.
viride TS strain. However, its industrial production has not
yet attracted much attention. Considering this fact, it could
also be used as flavoring agent particularly for cheese and
fruity flavors (Patton 1950; Hagedorn and Kaphammer 1994).
Use of these secondary metabolites in food industry has
become a major thrust area, making Trichoderma a versatile
fungus with application in natural as well as in industrial Fig. 3 Relative advantages of using formulation of antimicrobial sec-
systems. ondary metabolites in comparison to whole organism application
540 Appl Microbiol Biotechnol (2014) 98:533–544

growth promoting fungus (PGPF) and biological control po- Balde ES, Andolfi A, Bruyere C, Cimmino A, Theys DL, Vurro M,
Damme MV, Altomare C, Mathieu V, Kiss R, Evidente A (2010)
tential. But these properties are fully expressed in field only
Investigations of fungal secondary metabolites with potential anti-
when, firstly, an area-specific strain is applied in the respective cancer activity. J Nat Prod 73:969–971
region and, secondly, when the Trichoderma spore formula- Baldwin JE, Adlington RM, Chondrogianni J, Edenborough MS,
tion used has retained ample viability (high spore count). On Keeping JW, Ziegler CB (1985) Structure and synthesis of new
cyclopentenyl isonitriles from Trichoderma hamatum (Bon.) Bain.
the contrary, a formulation of secondary metabolites would
aggr. HLX 1379. J Chem Soc Chem Commun 12:816–817
have a much longer shelf life, a higher efficiency against soil Benitez T, Rincon AM, Limon MC, Codon AC (2004) Biocontrol mech-
borne phytopathogens particularly against bacteria; also, a anisms of Trichoderma strains. Int Microbiol 7:249–260
consortium of antimicrobial metabolites against individual Berg A, Wangun HVK, Nkengfack AE, Schlegel B (2004) Lignoren, a
new sesquiterpenoid metabolite from Trichoderma lignorum HKI
pathogens could be formulated and used regardless of geo-
0257. J Basic Microbiol 44:317–319
graphic location where the incidence of that particular disease Berkaew P, Soonthornchareonnon N, Salasawadee K, Chanthaket R,
is high. This approach would be unsurpassed by current Isaka M (2008) Aurocitrin and related polyketide metabolites from
technology, as here the formulation would specifically target the wood-decay fungus Hypocrea sp. J Nat Prod 71:902–904
Blight MM, Grove JF (1986) Viridin. Part 8. Structures of the analogs
a particular pathogen while remaining soil microbiota would
virone and wortmannolone. J Chem Soc Perkin Trans 1:1317–1322
remain unaffected (Fig. 3). Thus, further studies on the devel- Boyd RK, McAlees AJ, Taylor A, Walter JA (1991) Isolation of new
opment of pathogen-specific secondary metabolite-based for- isocyanide metabolites of Trichoderma hamatum as their (η5-
mulations are urgently needed. pentamethylcyclopentadienyl) or (μ5-
ethyltetramethylcyclopentadienyl) bis (lthiocyanato) rhodium com-
plexes. J Chem Soc Perkin Trans 1:1461–1465
Acknowledgments Chetan Keswani is grateful to Banaras Hindu Uni-
Brewer D, Gabe EJ, Hanson AW, Taylor A, Keeping JW, Thaller V, Das
versity, Varanasi, for providing CRET-UGC fellowship.
BC (1979) Isonitrile acids from cultures of the fungus Trichoderma
hamatum (Bon.) Bain. aggr., x-ray structure. J Chem Soc Chem
Commun 23:1061–1062
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