Verma 2007

Biochemical Engineering Journal 37 (2007) 1–20
Review
Antagonistic fungi, Trichoderma spp.: Panoply of biological control

Mausam Verma a , Satinder K. Brar a , R.D. Tyagi a,∗ , R.Y. Surampalli b , J.R. Valéro a
a INRS-ETE, Université du Québec, 490, Rue de la Couronne, Québec, Canada G1K 9A9
b US EPA, P.O. Box-17-2141, Kansas City, Kansas, KS 66117, United States
Received 31 October 2005; received in revised form 6 May 2007; accepted 18 May 2007
Abstract
Trichoderma spp. have been widely used as antagonistic fungal agents against several pests as well as plant growth enhancers. Faster metabolic
rates, anti-microbial metabolites, and physiological conformation are key factors which chiefly contribute to antagonism of these fungi. Myco-
parasitism, spatial and nutrient competition, antibiosis by enzymes and secondary metabolites, and induction of plant defence system are typical
biocontrol actions of these fungi. On the other hand, Trichoderma spp. have also been used in a wide range of commercial enzyme productions,
namely, cellulases, hemicellulases, proteases, and ␤-1,3-glucanase. Information on the classification of the genus, Trichoderma, mechanisms of
antagonism and role in plant growth promotion has been well documented. However, fast paced current research in this field should be carefully
updated for the fool-proof commercialization of the fungi. The aim of this review is to sum up the BCA activity potential of these fungi and to
shed light on commercial production processes. In this regard, this review focuses on Trichoderma spp. discussing different aspects—pest control,
growth promotion, bioremediation, production processes and market values. Nevertheless, more research and review of the information regarding
these biocontrol agents are needed to exploit their actual potential, which is the salient objective of this review.
© 2007 Elsevier B.V. All rights reserved.
Keywords: Antagonism; Biocontrol agents; Microbial propagules; Trichoderma spp.; Wastewater; Wastewater sludge
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.1. Existence of biological control agents (BCAs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.2. Fungal BCAs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.3. Status of Trichoderma spp. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
1.4. Constraints in commercialization of Trichoderma spp. BCAs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2. Pest control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1. Fungi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1.1. Trichoderma spp. as biofungicides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1.2. Modes of action . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1.3. Application in wood preservation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.1.4. Application in agriculture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
2.2. Bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.2.1. Limited application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.2.2. Potential—future application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.3. Invertebrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.3.1. Application potential . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.3.2. Rhizosphere—nematodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.3.3. Foliar application—aphids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
2.4. Weeds. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
∗ Corresponding author.
E-mail address: [email protected] (R.D. Tyagi).
1369-703X/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.bej.2007.05.012
2 M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20
3. Growth promotion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.1. Direct effect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.1.1. Metabolite production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.1.2. Participation with ectomycorrhizal sphere in growth promotion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.2. Indirect effect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.2.1. Induction of plant defence mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
4. Bioremediation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
5. Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1. Microbial propagules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.1. Spore/conidia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.1.2. Mycelium and chlamydospore . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2. Mass scale production strategies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2.1. Solid-state fermentation (SSF), and liquid fermentation (LF) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.2.2. Combined process. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.3. Solutions to improve LF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.3.1. Inoculum effect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.3.2. C:N ratio and level . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
5.3.3. Nature of C and N . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.3.4. Sporulation inducer compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.3.5. Physiochemical production parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.3.6. Fermentation modes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.4. Selection of raw materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.4.1. Conventional/semi-synthetic substrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.4.2. Alternate/recycled substrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.4.3. Wastewater and wastewater sludge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
5.5. Standard bioassay for quality control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
5.6. Trichoderma spp. based formulations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
5.7. Integrated pest management with Trichoderma spp. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
6. Market potential . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
7. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
1. Introduction trum in terms of disease control and production yield [5]. In

this context, Trichoderma spp. have been the cynosure of many
1.1. Existence of biological control agents (BCAs) researchers who have been contributing to biological control
pursuit through use of fungi [6–17]. Furthermore, Trichoderma
Phytopathogenic microorganisms and insects have been co- spp. share almost 50% of fungal BCAs market, mostly as
existing with plants since the very beginning of agricultural soil/growth enhancers and this makes them interesting candi-
evolution. Despite being a natural phenomenon, their mutual dates to investigate [4].
existence has adversely affected agriculture and forests from
time to time. With the advent of technology, physico-chemical 1.3. Status of Trichoderma spp.
methods have been adopted to mitigate the phytopathogenic
impacts on agriculture and forests. Use of crude (ash, raw extract Although, Trichoderma spp. have been known for a long
of certain plants, lime) and chemical pesticides, physical traps time (since 1865 [18]), the taxonomy and species identifica-
for insects [1] are examples of these control methods. How- tion were vague until around 1969 [19]. In fact, Druzhinina and
ever, later on, integration of biological control with pre-existing Kubicek [20] have extensively reviewed species concepts and
methods has further revolutionized the agricultural and forest biodiversity in Trichoderma fungi. The authors have mentioned
pest management. that Trichoderma fungi are difficult to distinct morphologically,
however, the phylogenetic classification has rapidly reached
1.2. Fungal BCAs 100 [21], and it is expected to increase consistently. In this
context, the advancements as well as limitations of modern
Currently, the role of BCAs is a well established fact and methods like genealogical concordance phylogenetic species
has become increasingly crucial, and in several cases, com- recognition (GCPSR) and DNA-barcode system for safe iden-
plementary or even replacing the chemical counterparts where tification of Trichoderma spp. warrant future investigations.
antagonistic fungi play an important part [2–4]. Fungal based The GCPSR requires the analysis of trees of several unlinked
BCAs have gained wide acceptance next to bacteria (mainly, genes, whereas, DNA-barcode system is based on the defined
Bacillus thuringiensis), primarily because of their broader spec- nucleotide sequence differences of different Trichoderma spp.
M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20 3
Nevertheless, application of Trichoderma spp. as BCAs in the cesses. In this regard, this review focuses on Trichoderma spp.
environment as well as the reported epidemics of commercially discussing different aspects—pest control, growth promotion,
grown mushroom (Agaricus bisporus) [22] and harmful effects bioremediation, production processes and market values.
on immunocompromised mammals [23] are reasons which war-
rant efficient and reliable species identification for Trichoderma 2. Pest control
fungi.
The widespread application of Trichoderma spp. as BCAs has 2.1. Fungi
been exploited and reported only lately against several soil-borne
phytopathogenic fungi [24,25]. Akin to most fungal BCAs, Tri- 2.1.1. Trichoderma spp. as biofungicides
choderma spp. can be efficiently used as spores (especially, In general, the current literature indicates that Trichoderma
conidia), which are more tolerant to adverse environmental con- spp. have been mostly used as biofungicide agents (Table 1).
ditions during product formulation and field use, in contrast to First report on the subject may be credited to Coley-Smith
their mycelial and chlamydospore forms as microbial propag- et al. [34] who by means of microtome sections have shown
ules [26]. Nevertheless, the presence of a mycelial mass is also that medulla of infected sclerotia of Sclerotium delphinii were
a key component for the production of antagonistic metabolites completely replaced by hyphae and chlamydospores of Tricho-
[7,27]. Conidia and mycelia can be produced in either solid-state derma hamatum on agar plates. Likewise, Henis et al. reported
or liquid fermentation. In general, liquid fermentation is more mycoparasitism (penetration and infection) of Trichoderma spp.
suitable method over solid-state fermentation for large scale against Sclerotium rolfsii, where chlamydospores were abun-
production, still special techniques are required for abundant dantly produced in contrast to conidia within the infected fungal
conidia production. sclerotia [35].
Trichoderma fungi are well known for their antagonism
against several soil-phytopathogens, involving fungi, inverte- 2.1.2. Modes of action
brates, and bacteria (Table 1). Their BCA activity is mainly 2.1.2.1. Mycoparasitism. According to Punja and Utkhede
attributable to various anti-microbial/antagonistic compounds [36], Trichoderma spp. are the most widely studied myco-
they produce, in addition to their aggressive mode of growth parasitic fungi. However, their mycoparasitism is difficult to
and physiology. Full exploitation of the BCA potential of Tri- demonstrate in situ until very recently due to technical dif-
choderma spp. could easily provide growth enhancement of ficulties in making in situ microscopic observations (e.g.,
domestic plants, green house plants, and agricultural crops. fluorescence imaging and differential staining), such as at the
soil–root interface. Moreover, techniques involving antibodies,
1.4. Constraints in commercialization of Trichoderma spp. such as combined baiting-ELISA (enzyme linked immunosor-
BCAs bant assay) techniques to detect Trichoderma spp. in composts,
would certainly increase our understanding of the mycoparasitic
Trichoderma spp. are also preferred in bioremediation due interaction of these fungi [37].
to the production of metabolites that are rich in peroxidases, Previously, Cook [38] classified the mycoparasitic interac-
and laccase enzymes [28,29]. Despite all the acquired under- tions as: (1) replacement (unilateral antagonism), (2) deadlock
standing about antagonistic action and growth promotion of (mutual antagonism), and (3) intermingling (no antagonism),
the Trichoderma spp., there are nevertheless some hurdles to with lack of explanation at microscopic level. However, more
their widespread success: (a) most of Trichoderma spp. based recently the understanding of mycoparasitism has consider-
BCAs are unregistered and are being marketed simply as “soil ably improved [7,14,27,39–44]. Interestingly, the studies were
enhancers”, probably due to lack of “well defined” modes of carried out at genetic [44,45] and microscopic [27,43] levels.
action of these fungi and their underdeveloped bioassay methods However, a broader concept concerning living plants (with the
(to ensure product quality) [4]. Furthermore, the registration of exception of preservation of wood, where Trichoderma spp.
Trichoderma spp. based BCAs as fungicides and growth promot- alone kill plant pathogenic fungi; discussed later), would be that
ers is time-consuming, expensive and frequently without well after being treated with mycoparasites, plants induce defence
defined protocols; (b) raw materials like, glucose, sucrose, corn mechanisms on their own. Further, this phenomenon leads to
steep liquor, wheat bran, soya meal, fish meal, used in culture production of fungal inhibitory compounds by plants in addi-
media for production of the fungi are very costly [30,31]; (c) tion to Trichoderma spp., thereby, facilitating mycoparasitism.
low efficacy; (d) low spore yield; (e) difficulties in quantifica- Moreover, the mycoparasitism shown by Trichoderma sp. was
tion of BCA activity. This has encouraged many researchers to host specific (e.g., Pythium oligandrum) [43]. Recently, the
investigate agricultural wastes [32], industrial wastes [13], and role of extracellular enzymes has been well documented by
municipal wastes [33] as probable substrates for Trichoderma several researchers (e.g., proteolytic enzymes [46,47]; ␤-1,3-
spp. production. glucanolytic system [48–50]; chitinase [44,51]). The complex
To recapitulate, the essential information regarding Tricho- group of extracellular enzymes have been reported to be a key
derma spp. as BCA is scarce as most of the current literature is factor in pathogen cell wall lysis during mycoparasitism.
focused mainly on its commercial enzyme production capacity.
The aim of this review is to sum up the BCA activity potential 2.1.2.2. Antibiosis. Antibiosis is the process of secretion of
of these fungi and to shed light on commercial production pro- anti-microbial compounds by antagonist fungi to suppress
Table 1
4
List of different Trichoderma spp. and respective BCA facts
Active agent Antagonist against Responsible metabolites/factors Disease/epidemic control
Fungi
T. harzianum 1051, T. harzianum 39.1 Crinipellis perniciosa Chitinase, N-acetylglucosaminidase, ␤-1,3-glucanase, Witches’ broom disease (Crinipellis perniciosus) of
total cellulase, endoglucanase, aryl-␤-glucosidase, Cocoa [63]
␤-glucosidase, protease and amylase
T. lignorum, T. virens, T. hamatum, T. Rhizoctonia solani Unknown inhibitory substances; extracellular Damping-off of bean [8,142,160,161]
harzianum and T. pseudokoningii metabolites or antibiotics, or lytic enzyme action
(Rifai)
T. viride, T. harzianum Aspergillus flavus and Fusarium moniliforme Lipolytic, proteolytic, pectinolytic and cellulolytic Fungal—seed-associated [10,11]
enzymes. Unknown (mycotoxins) antibiotic
compounds (e.g., peptides, cyclic polypeptides)
T. harzianum, BAFC 742 Sclerotinia sclerotiorum, BAFC 2232 ␤-1,3-Glucanase and chitinase Fungal—soybean plant [55]
T. sp. Sclerotium rolfsii Competitive inhibition Rotting of common vegetables [162]
M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20

T. harzianum 25, T. viride Serpula lacrymans Antibiotic; anthraquinones Fungal wood decay [70]
T. harzianum Alternaria alternata Endo-chitinase Fungal plant disease [140]
T. virens “Q” strain Rhizopus oryzae/Pythium sp. Plant phytoalexin induction by antibiotic compound, Cotton seedling disease [69,163,164]
gliovirin
T. viride isolate, T60 Soft rot and basidiomycetes decay fungi, namely, Volatile organic compounds, lytic enzymes and Sap stain discoloration of pine and spruce trees
Coniophora puteana, Postia placenta and Serpula soluble antibiotics; nutrient competition [165,166]
lacrymans
T. virens isolates GL3 and GL21; T. Rhizoctonia solani, Pythium ultimum, and Antibiotics gliovirin and gliotoxin, and other Damping-off of cucumber [14,65,167]
harzianum T-203 Meloidogyne incognita inhibitory metabolites
T. harzianum, T. aureoviride, T. koningii Pyrenophora tritici-repentis (Died) Drechs. Lytic enzymes such as chitinases and proteases; Tan spot and leaf blotch of wheat [85,168,169]
(anamorph = Drechslera tritici-repentis (Died) antibiotics; mycoparasitism
Shoem.
T. viride Colletotrichum truncatum Volatile compounds and non-volatile antibiotics, Brown blotch disease of cowpea [84]
viridin with anti-fungal and anti-bacterial properties
T. aureoviride T122, T. harzianum T66 Fusarium, Pythium, and Rhizoctonia strains ␤-Glucosidase, cellobiohydrolase; ␤-xylosidase and General fungal plant disease [108,170–172]
and T334, and T. viride T124 and T228 protease enzymes
T. viride, T. pseudokoningii and T. koningii Sclerotium cepivorum Volatile organic compounds, lytic enzymes and Allium (onion seedlings) white rot [141,173,174]
soluble antibiotics
T. harzianum Fusarium udum Lytic enzymes such as chitinases and proteases; Pigeonpea wilt [175,176]
antibiotics
T. harzianum Penicillium expansum Enzymes; antibiotics; mycoparastitism Apple blue and gray mold [80,81]
T. virens and T. harzianum Rhizoctonia solani Enzymes; antibiotics Stem canker or black scurf of potato [64]
T. harzianum, T. koningii Fusarium culmorum, Botrytis cinerea and Anti-fungal compound: 6-n-pentyl-2H-pyran-2-one Plant pathogens [41,47,83]
Rhizoctonia solani (6PAP); lytic enzymes, mainly, chitinases,
␤-1,3-glucanase, and proteases
T. koningii, T. aureoviride, T. Sclerotinia sclerotiorum Antagonism—enzymes; antibiotics and Sunflower head rot [86]
longibrachiatum mycoparasitism
T. harzianum Alternaria alternata Fungal cell wall degrading enzymes Fungal infection of leaves, stems, flowers and fruits
of annual plants, especially vegetables and
ornamentals [140]
T. harzianum and T. viride Lasiodiplodia theobroma; Diplodia natalensis; Anti-fungal metabolites; non-volatile and volatile Fruit rot/wilt of banana; mango stem-end rot and
Botryodiplodia theobromae; Fusarium moniliforme antibiotics post-harvest rotting of yams [78,79,177,178]
var subglutinans; Penicillium oxalicum Currie;
Penicillium sclerotienum Yamamoto; Aspergillus
niger van Tiegh; Aspergillus tamarii Kita;
Rhizoctonia sp.
T. asperellum Fusarium oxysporum Antibiosis, mycoparasitism and competition for Wilt of tomato [179]
nutrients
T. harzianum Aphanomyces cochlioides, Rhizoctonia solani, Enzymes; antibiotics and anti-fungal properties Common plant fungal diseases [180]
Phoma betae, Acremonium cucurbitacearum, and
Fusarium oxysporum f. sp. radicis-lycopersici
T. harzianum Botrytis cinerea and Mucor piriformis Antagonistic factors like enzymes and antibiotics Post-harvest rotting of strawberries [82]
Bacteria
T. viride Anaerobic bacteria—Bacteroides fragilis Heptelidic acid—antibiotic action [90,91]
T. hamatum Rumen bacteria—Escherichia coli, Megasphaera Antibiotic activity of an isocyanide Control of rumen bacteria [89]
elsdenii, Streptococcus bovis, Bacteroides metabolite—3-(3-isocyanocyclopent-2-enylidene)
ruminicola, B. succinogenes, Succinivibrio propionic acid
dextrinosolvens, Ruminococcus albus, R.
flavefaciens
T. viride Serratia sp. Non-volatile and volatile antibiotics Post-harvest rotting of yams [177]
T. harzianum (Rootshield® ) Clavibacter michiganensis Unknown Bacterial canker [87]
M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20

T. virens, T. koningii Bacillus subtilis Gliotoxin, dimethylgliotoxin, heptelidic acid, viridiol Biocontrol of plant diseases [67]
and viridin
Invertebrates (insects and nematodes)
T. viride, T. koningii, T. longibrachiatum, Atta cephalotesa (a leaf-cutting, fungus growing ant) 1,3-␤-Glucanase, chitinases, proteases, and lipases. Damage to plant leaves by ants [15,181]
T. hamatum, T. harzianum Unknown antibiotic compounds
T. harzianum Rifai ITEM 908 and ITEM Schizaphis graminum (aphid) Polysaccharide lyases, proteases, and lipases Poisoning of cereal crops
910
Trichoderma spp. Caenorhabditis elegans; Meloidogyne incognita Enzymes; antibiotics Root-knot nematode of horticultural crops [100]
Plants (weeds)
T. virens (=Gliocladium virens) Setaria viridis and Amaranthus retroflexus Phytotoxins, including viridiol, gliovirin, gliotoxin, Broadleaf and grass weeds [16,17,66,163]
and viridian; allelochemicals
T. viride Lantana camara; Rottboellia cochinchinensis; Phytotoxins and lytic enzymes Invasive weed [102]
Mikania micrantha
T. harzianum Phytophthora capsici; Phytophthora erythroseptica Antibiosis; enzymes Rotting of Capsicum annuum roots [155,182,183]
T. harzianum Verticillium dahliae Unknown Potato disease control [184,185]
T. koningii Gaeumannomyces graminis var. tritici Anti-fungal compounds Protect wheat against take-all disease [186,187]
T. virens V. dahliae Induction of terpenoids Verticillium wilt of cotton [185]
a Indirect antagonism: Trichoderma spp. inhibits the symbiotic fungus of Atta cephalotes, thereby, affecting the normal growth of the insect.
5
and/or kill pathogenic fungi in the vicinity of its growth area in vitro tests, yet their in situ action was inefficacious as observed
[52]. earlier [72]. However, further studies by Bruce et al. [73] showed
Menendez and Godeas [53] reported a biocontrol study of that in situ BCA potency of Trichoderma spp. was still effec-
Trichoderma harzianum against Sclerotinia sclerotiorum—a tive. In cases of lesser efficacy, a non-uniform distribution or
soil-borne plant pathogen attacking many economically impor- impoverishment of Trichoderma spp. and compartmentalization
tant crops, such as, soybean. The authors studied antibiosis of pathogenic fungi could be involved. Therefore, suitable in
of T. harzianum against the plant pathogen, assuming that the situ application techniques should be of prime importance, for
beneficial effect was due to concurrent mycoparasitism and com- a successful field application of BCAs.
petition as reported earlier in a similar study [54–56]. Meanwhile, Score and Palfreyman [70] postulated that the
In another example, despite close contact between hyphae presence of varying nutrient concentrations in wood might have
of Trichoderma spp. and Fusarium moniliforme/Aspergillus a marked effect on the nature of antagonistic interactions. They
flavus on co-culturing, hyphal penetration was absent, sug- also showed that varying nutrients in enriched media produced a
gesting that mycoparasitism was not the sole cause for the clear effect on hyphal extension rate of several Trichoderma spp.
observed inhibitory effects [11]. Therefore, metabolites pro- affecting their BCA activity. However, determination of actual
duced by Trichoderma spp. (e.g. volatiles, extracellular enzymes BCA activity would require extensive studies on wood like, the
and/or antibiotics) were considered to be the probable ele- possible interactions of various Trichoderma spp. metabolites
ments involved in antibiosis. Trichoderma spp. have been also with major wood components, namely, tannin, and lignin.
effective in cases of a wide host range and in hindering the
longevity of sclerotia of pathogenic fungi. Recently, Szekeres 2.1.3.2. Cosmetic woods. Ejechi [12] investigated the ability of
et al. [57] have reviewed antagonistic metabolites produced Trichoderma viride to inhibit the decay of obeche (Triplochiton
by Trichoderma spp. The metabolites are linear, amphipathic sceleroxylon) wood by the decay fungi Gloeophyllum sp. and G.
polypeptides, namely, peptaibols and peptaibiotics. They also sepiarium under field conditions. The study was conducted over
discussed the physico-chemical and biological properties of a period of 11 months, covering dry and wet season in tropi-
these antibiotic compounds which included the disruption of cal environment. T. viride exhibited total inhibition of the decay
lipid membranes, anti-microbial activities, and induction of fungi by means of mycoparasitism and competition for nutrients.
plant resistance. Trichoderma spp. are currently the most extensively investi-
gated biocontrol fungi for forest product preservation, and, on
2.1.2.3. Competition. Celar [58] conducted a study on the a number of occasions, have successfully provided more effec-
forms of nutrients commonly available to phytopathogenic and tive protection against certain wood decay fungi in comparison
antagonistic fungi. Earlier study reconfirmed the findings of to other antagonistic fungi, e.g., Penicillium sp. [71,74]. Using
Blakeman [59] that shortage of easily accessible nutrients for modified versions of American [75] and European [76] standard
microorganisms, especially of those living in soil and on plant test methods as well as a soil burial test system, Tucker et al. [77]
surfaces, could result in explicit nutrient competition among have shown that certain isolates of Trichoderma spp. were totally
microorganisms [60,61]. In addition, Trichoderma spp. could effective in protecting wood against certain basidiomycetes.
compete and sequester ions of iron (the ions are essential for the
plant pathogen, Serpula lacrymans as part of a non-enzymatic 2.1.4. Application in agriculture
complex) by releasing compounds known as siderophores [62]. 2.1.4.1. Fruits and vegetables. Several Trichoderma spp. have
Thus, the cited examples confirmed that significance of compe- also been used to protect commercially important fruits and
tition for nutrients between Trichoderma and pathogenic fungi. vegetables, such as banana, apple, strawberries, mango, potato,
Several authors have highlighted the significance of lytic and tomato during post-harvest storage (Table 1). Mortuza and
enzymes in BCA activity and studied isolates of Tricho- Ilag [78] utilized 10 isolates of Trichoderma spp. including T.
derma spp. with cellulose and chitin degradation characteristics harzianum and T. viride against the banana fruit rot pathogen,
[63–65]. Hutchinson [66] and Hanson and Howell [67] have Lasiodiplodia theobromae. Their investigation confirmed that
reported the significance of secondary metabolites (antibiotic mycoparasitism was associated with antibiosis and competition
activity) in antagonistic action of Trichoderma spp. against for substrate. The authors compared the cultural filtrates of Tri-
pathogenic fungi Pythium ultimum and Rhizoctonia solani. choderma spp. with a chemical fungicide, namely, BenomylTM ,
However, there seems to be a general consent on the combined and concluded that Trichoderma spp. based fungicide could not
synergistic effect of the two factors (enzymes and antibiotic be totally as effective as their chemical counterparts. However,
compounds) [52,68,69]. the authors underestimated the role of Trichoderma spp. based
formulations for field application, which might substantially
2.1.3. Application in wood preservation enhance the BCA activity.
2.1.3.1. Hardwood. Biological control studies to protect Biological control of mango stem-end rot using T. viride was
wooden distribution poles by Trichoderma spp. have also been studied by Moreno and Paningbatan [79], where they reported
carried out against the dry rot fungus, S. lacrymans [70] and mycoparasitism and antagonism as major BCA activity factors.
brown rot fungi, namely, Antrodia carbonica and Neolentinus Batta [80,81] examined the invert-emulsion formulation of T.
lepideus [71]. From these studies, it was concluded that although harzianum Rifai against blue mold infection of apple to control
Trichoderma spp. displayed a killing action against these fungi in post-harvest fruit decay. The author attributed the BCA activity
Fig. 1. Growth promotional activities of Trichoderma spp. Indirect: (a) mycoparasitism, (b) competition; direct: (c) mycelial growth around plant rhizosphere and
production of metabolites.
of Trichoderma spp. to the necessary period of conidial humecta- based on their mycoparasitic and antibiotic activities. However,
tion in order to germinate and penetrate into the plant pathogenic particular factors triggering antagonism (molecular signal for
fungi. It was also reported that the invert-emulsion was better mycoparasitism, as Trichoderma spp. were able to recognize
application mode for fungal BCAs like Trichoderma spp. Brewer their specific host) were not available. In another instance, the
and Larkin [64] reported Trichoderma spp. to be potential antag- beneficial action of T. virens for pre-treatment of cotton seedlings
onists for stem disease of potato in comparison to several other has been reported. It was demonstrated that the induction of
fungal BCAs. On the other hand, Hjeljord et al. [82] reported plant defence system and the suppression of pathogen germina-
that application of Trichoderma spp. on greenhouse strawber- tion by antagonistic compounds produced by germinating cotton
ries could control post-harvest rotting. They emphasized that seedlings were the dominant biocontrol mechanisms [69].
temperature and nutrients influenced the BCA activity of T. Thus, it is evident that the processes of mycoparasitism,
harzianum. Results of Cooney and Lauren [83] confirmed antag- antibiosis, and competition for substrates are well documented,
onistic metabolite production by a Trichoderma sp. induced by as exemplified in Fig. 1. However, the exact phenomena involved
the presence of pathogenic fungi (300–700% increase in metabo- in the overall antagonistic action of Trichoderma spp. is still not
lite production in the presence of plant pathogenic fungi). well understood. It is postulated that Trichoderma spp. initially
succeed in antagonism via hyphal interactions, probable primal
2.1.4.2. Crops/seeds. Trichoderma spp. are well-recognized step in antagonism. Later, the antagonistic fungi kill the phy-
fungal antagonists of crops/seeds pathogens. Biocontrol of topathogenic fungi by means of toxins and consume them using
brown blotch of cowpea caused by Colletotrichum truncatum a combination of lysozymes [4].
by pre-treatment of cowpea seeds in T. viride spore suspension On the other hand, the fungicidal activity of Trichoderma spp.
was considered to be due to both mycoparasitism and antibiosis is well known with respect to most of the fungal phytopathogens.
[84]. Similarly, the application of Trichoderma spp. for the con- Most researchers agree to the fact that the antagonists (Tri-
trol of wheat [85] and sunflower [86] fungal diseases has been choderma spp.) control pathogens via interlinked synergistic
complex strategies of mycoparasitism, antibiosis, and compe- nematodes were mainly antagonized by parasitism and antibiosis
tition. Therefore, in order to exploit maximum potential of akin to fungal pathogens.
Trichoderma spp. against fungal pathogens, factors (e.g., distri-
bution of inoculum at infected sites, concentration of inoculum, 2.3.3. Foliar application—aphids
specificity towards pathogens, environmental conditions and The insecticidal activity of two strains of T. harzianum against
enrichment medium—if any) affecting one or many of the antag- aphids has been reported [101]. Since T. harzianum is currently
onistic strategies should be optimized. considered as an important BCA candidate, its insecticidal activ-
ity has further importance. The authors proposed that the action
2.2. Bacteria of toxins produced by T. harzianum strains was facilitated by
cuticle degrading extracellular enzymes (proteases and chiti-
2.2.1. Limited application nases) that enabled the insertion of the toxins inside cuticle.
In contrast to other fungi, Trichoderma spp. have been Thus, many Trichoderma spp. are reported as occasional par-
reported to have limited applications in biocontrol of pathogenic asites of invertebrates. Therefore, the actual potential of these
bacteria. An immediate explanation would be that bacteria gen- fungi in the natural suppression of insects affecting economical
erally have a faster metabolic rate than fungi. Thus, antagonism crops is still underrated. This furthermore warrants research on
via physical interaction such as, mycoparasitism would be too using Trichoderma spp. based BCAs to cope with insect related
slow to be effective from BCA point of view, where faster action plant disorders.
is a must. However, if the formulated metabolites from Tricho-
derma spp. were considered, the BCA potential of antagonist 2.4. Weeds
fungi would be considerably higher.
Herbicidal action of Trichoderma spp. has also been reported
2.2.2. Potential—future application by some authors ([16,17,66,102]; Table 1), however, weed con-
In view of limited studies on anti-bacterial action of Tri- trol by using Trichoderma spp. is still a relatively unexplored
choderma spp., very few examples have been reported in the field. Heraux et al. [16,17] have reported the weed control action
literature, which have been listed in Table 1. Altogether, anti- of T. virens. The study reported that T. virens with and without
bacterial action of Trichoderma spp. was based only on the action combination of composted manure and rye cover crop controlled
of antibiotic compounds produced and no physical interaction a mixed community of grass weeds and broadleaf weeds. The
between antagonist and pathogen was mentioned [67,87–91]. weed control action was comparable to chemical herbicides,
Therefore, in the present scenario, despite Trichoderma spp. namely, metribuzin, sethoxydim and ethalfluralin. T. virens com-
possessing anti-bacterial potential, their applicability cannot be posted chicken manure and rye cover crop has showed herbicidal
advocated and actually employed in field set-ups. activity [16,17,66]. The herbicidal activity was attributed to: (a)
viridiol produced by T. virens and (b) the herbicidal molecules,
2.3. Invertebrates namely, (3H)-benzoxazolinone (BOA) and 2,4-dihydroxy-1,4-
(2H)-benzoxazine-3-one (DIBOA) released during composting
2.3.1. Application potential of chicken manure and rye cover crop [16,17]. However, there
Trichoderma spp. are basically soil-borne saprophytic fungi is a need for future studies to ensure reliable performance prior
which possess a symbiotic relationship with the plant rhizo- to commercial utilization of T. virens. In fact, utilization of Tri-
sphere [92]. In addition, they have innate ability to produce choderma as herbicides is a relatively new approach and is not
several chitin (a major component of cellular structure in inver- feasible under present scenario. Nevertheless, future research
tebrates) degrading enzymes (endochitinases and exochitinases) should be carried out to increase the concentration of viridiol
in order to survive on and/or antagonize pathogen organism and potential phytotoxins (Table 1) produced during fermenta-
[49,51,63,93]. tion. Agricultural residues other than rye cover crop and chicken
manure should also be examined in order to make the process
2.3.2. Rhizosphere—nematodes economically feasible.
Nematodes are mostly present in rhizosphere than in the bulk
soil, therefore, their antagonist(s) should also be in sync with 3. Growth promotion
rhizosphere. Fungi like Trichoderma spp. fit very well into this
category [94–100]. 3.1. Direct effect
Most studies on nematodes concurred that the promising fun-
gal antagonists—Trichoderma spp., had different and in fact 3.1.1. Metabolite production
multiple modes of action. For example, Trichoderma virens Literature is replete with many studies on plant growth
invaded, ramified, grooved and vacuolated the root-knot nema- promotional activity of Trichoderma spp. [4,36,42,103]. Some
tode eggs. Eapen et al. [100] reported easy staining of eggs for researchers have suggested production of growth hormones
microscopy due to the increased permeability of eggshell. The [104,105] and enhanced transfer of minerals to rhizosphere [104]
antagonistic action of Trichoderma spp. was chiefly attributed as direct factors governing spectacular performance of Tricho-
to chitinolytic activity of the fungi on cellular structure of nema- derma spp. based BCAs as also reported in Table 2. Besides,
todes, which is rich in chitin. Additionally, unlike bacteria, Trichoderma spp. have been reported to promote growth in
strawberries [106]. Despite all the research, there is a need for
Reference(s)
[188–190]
[106,191]
[194,195]
extensive optimization of application conditions and active field
[192]
[193]
[196]
studies.
[58]
3.1.2. Participation with ectomycorrhizal sphere in growth
promotion
Lindsey and Baker [107] demonstrated that the symbiosis of
Co-metabolism in the presence of glycerol

Co-metabolism of cyanide in the presence
Co-metabolism in the presence of glucose

mobilized N from organic compounds via
Mycorrhizal fixation of phosphorus and

T. viride in rhizosphere helped in growth of gnotobiotic (either,
Saprotrophic fungus, Trichoderma sp.
directly uptaking organic N from soil

Bioremediation/Suggested mode of
well defined or, no microflora present) tomato, thereby, showing

ectomycorrhizal fungi incapable of
growth promotional ability of this fungus. The physical pres-

ence of mycelial mass in rhizosphere in itself would serve as
Nutrient supplementation
appendage to the normal rhizosphere of plants, thereby, enhanc-
Competitive inhibition
ing nutrient uptake. Further, the advantages of Trichoderma spp.
growth promotion
based BCAs for growth promotion are enhanced due to symbio-

sis with ectomycorrhizal sphere [16,108,109]. Trichoderma spp.
of glucose
easily acquire nutrients from complex substrates like, protein-

nitrogen
tannin, and glucosamine, in soil due to their ectomycorrhizal

association. Further, the nutrients were easily utilized by the
plant due to their mutual symbiotic relationship. In particular,
Trichoderma spp. has been extensively utilized in waste com-
posting [17,66,109], which ultimately ends up in agricultural
Growth promoters/target compound
Rhizosphere enhancers—N and P

NH4 + –N; organic N (amino acid)
land, consequently affecting plant yield. The positive role of

Trichoderma spp. in ectomycorrhizal sphere has been also elab-
800–1100 mg PAHs/kg soil
orated by Wu et al. [110] which is an indirect mode for their

Nitrogen and phosphorus
NH4 + –N and NO3 − –N
plant growth promotional activity.

Cyanide to ammonia
3.2. Indirect effect

concentration
100 ␮g/g soil
3.2.1. Induction of plant defence mechanism

In addition to the well-recognized mycoparasitic nature of
Trichoderma fungi, induction of resistance against pathogens in
plants has also been reported [27,42,44,111] as indirect growth
promotion factor. The authors suggested that association of Tri-
Polyaromatic hydrocarbons (PAHs)
choderma with roots, and several lytic enzymes induced by plant

Bioremediation and suggested plant growth mechanisms by different Trichoderma spp.
Strawberry, Fragaria ananassa
defence system destroyed and consumed the pathogen cell wall.

Plant name/target compound
Finally, it provided nutrient to the plant as depicted in Fig. 1.

P. resinosa Ait (red pine)
A mycoparasitic interaction study in this context revealed that

ech42 (a chitinolytic enzyme encoding gene) transcription was
induced prior to physical contact of T. harzianum with host
pathogen fungus [44]. The authors also expressed the possibil-
Chickpea
Fluorene
Cyanide
ity of different mechanisms of induction for each type of lytic

Pine
enzyme and the necessity to characterize these enzymes and

their relevance in mechanisms of biocontrol.
Available literature demonstrates the plant growth promo-
tional activity of Trichoderma spp., still exact or quantitative
T. harzianum (O90, O77, O82 and O80) and
T. longibrachiatum, T. harzianum, T. viride,
BCA assessment is difficult due to multiple factors associated

T. hamatum, T. harzianum and T. koningii
and scarce information available. Nevertheless, there are many

one T. pseudokoningi strain (O10)
T. harzianum, T. viride and T. virens
Trichoderma spp. based commercial products in market which

aim at greenhouse plants (mainly ornamental and garden veg-
etables) as depicted in Table 3.
4. Bioremediation
T. harzianum DB11
and T. koningii
Trichoderma sp.
Trichoderma sp.
The concept of utilizing fungi for bioremediation of soil

contaminated with certain pollutants is relatively older. There
Table 2
is ample evidence of various Trichoderma spp. contribut-

T. sp.
ing to polycyclic aromatic hydrocarbons (PAHs) degradation,

whilst affecting native mycorrhizal fungi both positively and/or,
U.S., U.S., Belgium

Country registered
negatively [112]. Saraswathy and Hallberg [113] reported a max-
U.K., Sweden
imum of 75% removal for pyrene (4-ring PAH) at 50 mg l−1 for
Antagonism by enzymes and antibiotics New Zealand

U.S., Europe
axenic cultures of Trichoderma spp. while pyrene served as sole
Spain
Israel
Israel
India
carbon source.
U.S.
Katayama and Matsumura [28] demonstrated degradation

potential of rhizosphere-competent fungus Trichoderma sp.
against several synthetic dyes, pentachlorophenol, endosulfan,
Type of action as per manufacturer
Mycoparasite living on other fungi
and dichlorodiphenyl trichloroethane (DDT). Thus, Tricho-

derma spp. have found application in herbicide/pesticide laden
soil bioremediation as sustainable approach. Review of present
literature suggests that hydrolases, peroxidases, laccases and
Parasite, competitor
other lytic enzymes produced in abundance by Trichoderma

spp. are probable factors aiding in degradation of these con-
Mycoparasite
Antagonist
taminants.
From BCA point of view, bioremediation potential of fungi
like Trichoderma would be an additional advantage as this will
–
–
–
aid in soil enhancement where excessive use of herbicides should
rot, red rot, damping-off, Fusarium wilt on wide
spp., Fusarium spp., root rot, seedling rot, collar
Sclerotium rolfsii, Pythium spp., Fusarium spp.
Soil pathogens that cause damping off and root
Rhizoctonia solani, Sclerotium rolfsii, Pythium

For management of Rhizoctonia spp., Pythium
to be curtailed. However, this can be achieved, if and only if the

rot, esp. Rhizoctonia solani and Pythium spp.
Armillaria, Botryosphaeria and other fungal
contaminated soil is inoculated with Trichoderma spp. at regu-

Soil pathogens—Pythium, Rhizoctonia,
lar defined intervals. In fact, inclusion of Trichoderma spp. in

Growth promoter, Rhizoctonia solani,
Verticillium, Sclerotium, and others
“integrated pest management” program is a must. In the current

context, the scientific community believes that use of herbicides
cannot be significantly reduced due to immediate concern of
Botrytis cinerea and others
on nursery and field crops
needs of agricultural commodities.

Tree-bound pathogens
Therefore, consistent simultaneous application of some

“detoxifying” agents along with (Trichoderma spp. versus harm-
Pests controlled
variety of crops
ful pesticides/herbicides; the former can tolerate and degrade

application concentrations of several pesticides/herbicides)
diseases
would provide an agreeable soil environment. This will assure

not only the health of soil and plant, but also a sustained
crop yield protection. Future research must be streamlined
BioWorks, Wilbur-Ellis, Borregaard
J.H. Biotech, Mycontrol, Ltd., De
on consistent-simultaneous use of Trichoderma spp. as BCA

cum soil remediation agent along with obligatory pesticides/
Manufacturers and suppliers
herbicides.
Mycontrol (EfA1) Ltd.
Agrimm Technologies
5. Production
Bio-Innovation
Ecosense Labs
Makhteshim
5.1. Microbial propagules

Ceuster
Certis
[180]
List of Trichoderma fungi based BCAs and respective suppliers
5.1.1. Spore/conidia
The ultimate objective of any BCA lies in its feasibility of
RootShieldTM , BioTrek 22GTM ,
Source: http://www.google.com internet search engine.

TrichodowelsTM , TrichosealTM
economical mass production which also holds true for Tricho-

PromotTM , Trichoderma 2000,
derma spp. based BCAs. Further, from Table 3 and Fig. 2, it is

TrichopelTM , TrichojetTM ,
SupresivitTM , T-22GTM ,
obvious that Trichoderma spp. based BCAs are commercially

viable as numerous commercial products exist in market, still
12GTM
Trichoderma 2000
a majority of them (not presented here) are “unsung” BCAs. A

TrichodexTM
vast majority are rather being promoted as soil enhancer and/or

Trade name
T-22HBTM
Soil Guard
Biofungus
TUSAL®
BinabTM
growth promoter. Almost all available Trichoderma spp. based

Trieco
BCA products contain spores as active ingredients [4,36,80,81].

This could be attributed to the physiological aspects of their three
microbial propagules, namely, mycelia, conidia, and chlamy-
Gliocladium virens#
Beneficial organism
T. harzianum and T.
T. harzianum and T.
dospores [26,31,114]. The three propagules possess distinct

polysporum
physiological characteristics in terms of production, stability and

T. harzianum
T. harzianum
T. harzianum
T. harzianum
BCA activity. Therefore, it is imperative to select the best suit-

viride
T. viride
Table 3
T. spp.
able form of Trichoderma spp. propagules in order to efficiently

execute their BCA action.
aged major BCA producers to consider SSF as a viable mass

scale option [118].
Unfortunately, accurate information regarding factors direct-
ing sporulation process in LF of fungi, especially, for
Trichoderma spp. is scarce and incomplete, besides being pro-
prietary. Therefore, production of spores in LF still remains a
challenging task and warrants considerable research inputs.
5.2.2. Combined process

To overcome limitations of SSF and LF many researchers
also suggested hybrid strategies involving both SSF and LF.
Normally, LF is followed by SSF in many industrial produc-
tion processes. However, relevant information on production
techniques is so far evasive in current literature. In a typical Tri-
choderma spp. production process, 2–3 days old broth of LF is
used as inoculum for solid substrates, e.g., bran, rice, grain-husk
and others. The solid substrates thus inoculated were incubated
Fig. 2. Trichoderma spp. based biofungicide market statistics. Other biofungi-
cides include bacteria, nematodes and virus. Note: The market is based on
for further 2–8 days, followed by addition of formulation agents,
scattered data of registered biofungicides. e.g., carboxy methyl cellulose, silica, talc and moderate tempera-
ture (about 20–40 ◦ C) air-drying below 8–10% moisture content.
However, labour-intensive nature of similar techniques certainly
5.1.2. Mycelium and chlamydospore could not replace LF. Meanwhile, the problems in LF will no
Although, mycelia have excellent BCA activity, unfortu- doubt eventually be overcome.
nately, they cannot survive down stream processing steps such
as drying and hence are not useful [26]. On the other hand, 5.3. Solutions to improve LF
chlamydospores require a period of 2–3 weeks for cultivation
and likewise could not survive drying processes [31], albeit, 5.3.1. Inoculum effect
they are more stable than mycelia. Meanwhile, as said above, The ongoing research also indicates that several techniques
conidia are active as BCA, less susceptible to several environ- might help in improving sporulation during LF. For example,
mental conditions and could be produced faster (3–4 days) in fungal morphology and the general course of fermentation has
abundance [33]. Thus, production of Trichoderma spp. as coni- been primarily affected by the amount, type (spore or vegeta-
dia would be the best option from BCA application point of tive) and age of the inoculum [119–121]. In general, studies
view. However, the presence of mycelia along with conidia in carried out on fungi until now suggested that high spore con-
the production media cannot be ruled out. In addition, simul- centrations in inoculum (≤105 to 108 spores/ml) were not good
taneous mycelial production would insure presence of various for sporulation. Therefore, any direct role of inoculum (in form
essential metabolites (e.g., antibiotics) for BCA activity [66,67]. of spores) in sporulation could be ruled out. Nevertheless,
Thus, production of Trichoderma spp. containing conidia as same study also suggested that inoculum could be important
main propagules along with mycelia could be the best production for production of metabolite(s) and mycelial mass responsi-
strategy. ble for the BCA activity. Eventually, type, quantity, and age
of inoculum should be explored for probable enhancement of
5.2. Mass scale production strategies sporulation.
5.2.1. Solid-state fermentation (SSF), and liquid 5.3.2. C:N ratio and level
fermentation (LF) Variation in values of carbon: nitrogen (C:N) ratio in medium
Commercial success of Trichoderma spp. based BCAs would could also be helpful in sporulation/conidiation of Trichoderma
also require economically feasible mass scale production pro- spp. Olsson et al. [122] observed start-time of sporulation phase
cesses (as 35–40% costs of production depends on raw material). in Trichoderma reesei Rut-30 and remarked that decreasing
Of the two broad categories of production, namely, solid- C:N ratio adversely affected sporulation. Moreover, studies on
state fermentation (SSF) and liquid fermentation (LF), LF has other fungi, namely, Neurospora crassa [123] and Beauveria sp.
been adopted by many researchers despite lower sporulation [124] also showed marked effect of C:N ratio on sporulation.
[115,116]. Labour, scale-up, process control, productivity, mate- Therefore, manipulation of C:N ratio remains a viable option
rial handling (pumping, pressurized lines), compatibility with for further investigation on sporulation process of Trichoderma
pre-existing large scale facilities are some positive features of spp.
LF that encourage most researchers to pursue LF in lieu of SSF. As mentioned earlier, the antagonism study of Trichoderma
However, SSF is often preferred to LF, when production scale is spp. by Score and Palfreyman [70] used a standard complex
of moderate range and labour force is cheap [117]. Additionally, medium (malt extract agar, MEA) and a minimal essential
recent advancements in industrial automatization have encour- medium (MEM) designed to mimic the C:N ratio in the pathogen
infected wood. The findings revealed that Trichoderma spp. 5.4. Selection of raw materials
could be significantly antagonistic even at low nitrogen con-
tent. Therefore, any increase in C:N ratio owing to decrease in 5.4.1. Conventional/semi-synthetic substrates
N would not affect the BCA activity of these fungi. In other Several growth/production media for Trichoderma spp.
words, substrates with higher C:N ratio could be beneficial for spores production are presented in Table 4. Use of media
both sporulation and BCA activity. based on molasses, d-glucose, cellulose, or soluble starch
have been also reported by many authors [31,115,116]. In
5.3.3. Nature of C and N general, maximum sporulation of the order of 108 spores/ml
Sporulation of Trichoderma spp. is greatly affected by the fermentation broth could be achieved. However, it should be
nature of carbon and nitrogen sources used as substrates. Pas- noteworthy that production of Trichoderma spp. spores for
cual et al. [125] observed that peptone, free amino acids (e.g., BCA use (application in soil), using these substrates could not
arginine), mannose, xylose, and fructose induced high level of be economical, owing to high raw material cost and moder-
sporulations. It could be inferred that exploring various carbon ate sporulation. Meanwhile, a much higher spore concentration
and nitrogen sources and their pre-treatments could be potential (≈107 to 1013 CFU/ha application requirements will neces-
means to induce sporulation. sitate > 107 to 1010 CFU/g formulated product, consequently
requiring ≥107 to 1010 CFU/ml fermented medium) will be
required for field application [82].
5.3.4. Sporulation inducer compounds
Introduction of some triggering agents has also been help-
5.4.2. Alternate/recycled substrates
ful in sporulation process. These agents might be metal ions
In order to obtain crude proteins, enzymes as mycelial mass,
(e.g., manganese ions [126]), or complex organic compounds.
use of effluent of oil mill as substrate for producing Trichoderma
Roncal et al. [127] have isolated conidiogenol and conidio-
spp. has been investigated, where the fermentation time was
genone, tetracyclic diterpenes with potent and selective inducing
longer (≥2 weeks) [130] Felse and Panda [93,129] have reported
activity for conidiogenisis in Penicillium cyclopium. These
the use of untreated crab shell to increase chitinase production.
findings are still at their natal stage and might be helpful in
Olsson et al. [122] used cellulose, raw and treated sugar beet
practical application at later stage. Nevertheless, it appears
pulp without additional nutrients and Mendel’s medium compo-
that introduction of some metal ions and/or some complex
nents for the development of inoculum. Meanwhile, studies on
compounds to the growth medium could induce sporulation.
steam-pretreated willow (lignocellulosic material) [32,131] sug-
Perhaps, various metals and complex compounds in renewable
gested replacement of glucose and corn steep liquor in Tanaka
waste, could be helpful in achieving high spore concentra-
media (Table 4) by Avicel (10 g l−1 ), or by corn fibre dry mass
tion.
(20–140 g l−1 ). Several studies have been reported on the use of
alternative cheap raw material for Trichoderma spp. production
5.3.5. Physiochemical production parameters processes [132–135].
Environmental parameters like, temperature have been less Remarkably, in the preceding examples, the authors studied
critical parameters for sporulation [128]. Meanwhile, pH of cellulosic and hemicellulosic enzymes producer Trichoderma
the medium plays an important role. Although, conidial fungi spp. In all studies, the BCA potential of fermented products
can grow over a wide range of pH, they grow and sporulate was not the main objective of the authors. The sole presence
maximally near neutral pH [126]. Felse and Panda [129] inves- of metabolites in Trichoderma spp. based BCA is not sufficient
tigated the effect of agitation conditions on a Trichoderma sp. for its application viability, high spore concentration is equally
and inferred that extremely agitated conditions were not good important. In fact, most studies focused mainly on production of
for growth and sporulation. Dissolved oxygen and dissolved Trichoderma spp. for metabolites (enzymes and/or antibiotics)
carbon dioxide may also play an important role in inducing and not sporulation (an important factor for practical application
sporulation, probably by imposing mass transfer related stress of BCAs). Therefore, validation of commercial viability of these
conditions on the culture. The sporulation in fungi is also raw materials/processes from BCA point of view would require
an important mode of reproduction, therefore, sporulation of further investigations aimed at spore production and bioassay
Trichoderma spp. could also be correlated to the respiration against commercial pests.
quotient (RQ).
5.4.3. Wastewater and wastewater sludge
5.3.6. Fermentation modes Based on the preceding discussion for economical (low cost
In general, there are no research reports on the studies regard- raw material) and efficient production (higher sporulation) of
ing culture conditions like continuous and fed-batch culture Trichoderma spp. based BCAs, a novel array of substrates have
in fermenter that allow finer control of substrate concentra- to be explored. Akin to prior mentioned wastes, experiment-
tion (solids concentration, C:N ratio), biomass (mycelia and/or ing with wastewater and wastewater sludges (source of carbon,
spores concentration), single/multiple nutrient addition (feeding nitrogen, phosphorus, and other essential nutrients for many
strategy for complex and/or simple substrates). Thus, the above microbial processes) could provide probable viable solution to
listed parameters should be examined and studied adequately to combat the raw material cost and enhance sporulation, as in
induce sporulation. the case of B. thuringiensis [136]. In this context, Verma et
Table 4
Growth/production media used for the production of Trichoderma spp. for BCA use
Media Composition (g l−1 ) Remarks (spore yield) Reference
Basal salt medium Yeast extract, 0.5; (NH4 )2 SO4 , 1; MgSO4 ·7H2 O, [197]
Semi-synthetic (LF) [spore yield ≈ 106 to
0.3; KH2 PO4 , 1.36; either cellulose powder or the
107 ml−1 ], pHa 4–7, temperaturea : 28–30 ◦ C;
holocellulose, 10
incubation timea : 4–21 days
Mendel’s medium Glucose, 10; urea, 0.3; (NH4 )2 SO4 , 1.4; [30]
KH2 PO4 , 2.0; CaC12 ·2H2 O, 0.4; MgSO4 ·7H2 O,
0.3; peptone, 0.75; yeast extract, 0.25;
FeSO4 ·7H2 O, 0.005; MnSO4 ·4H2 O, 0.0016;
ZnSO4 ·7H2 O, 0.0014; CoC12 ·6H2 O, 0.020
Tanaka medium Glucose, 20; urea, 0.3; corn steep liquor, 10; [198]
(NH4 )2 SO4 , 1.4; KH2 PO4 , 2.0; CaC12 ·2H2 O,
0.4; MgSO4 ·7H2 O, 0.3; proteose peptone (Difco),
1; FeSO4 ·7H2 O, 0.005; MnSO4 ·4H2 O, 0.0016;
ZnSO4 ·7H2 O, 0.0016; CoC12 ·6H2 O, 0.004
M-CSL, SN, GT Glucose, 25; NaCl, 25; corn steep liquor, 5; [31]
molasses (blackstrap), 50. Sucrose, 20; NaNO3 ,
6; KH2 PO4 , 1.5; MgSO4 ·7H2 O, 0.5; CaCl2 , 20.
Glucose, 20; MgSO4 ·7H2 O, 1; KH2 PO4 , 2;
ammonium tartrate, 1; FeSO4 , 0.001
Corn cobs, wheat bran, cornmeal Quartz sand, 1000; corn cobs, 400 (in 1600 ml Complex (SSF) (liquid; any of, M-CSL, SN, [31]
liquid). Quartz sand, 1000; wheat bran, 400 (in GT, or water) [spore yield ≈ 107 to 108 g−1 ]
800 ml liquid). Quartz sand, 1000; cornmeal, 40
(in 200 ml liquid)
Czapek dox agar medium Sucrose, 20.0; NaNO3 , 2.0; MgSO4 ·7H2 O, 0.5; Synthetic (SSF) [134]
KCl, 0.5; FeSO4 , 0.01; agar, 20.0
PDA Potato starch; dextrose
Semi-synthetic (SSF)
Oat agar Oat flour, 30; agar, 15; ZnSO4 ·7H2 O, 0.01; [122]
CuSO4 ·5H2 O, 0.005
Botryosphaeran (exopolysaccharide (EPS)) Vogel minimal salts medium and 1.5 g l−1 of EPS Semi-synthetic (LF) [199]
as carbon substrate
Cranberry pomace-based medium Cranberry pomace, 10 g; CaCO3 , 0.5 g; water, Semi-synthetic (SSF), 5 mg/g pomace [13]
20 ml; NH4 NO3 , 0.5 g, and or fish protein protein
hydrolysate (FPH), 2 ml
Crude cell wall preparations from barley (In w/v%) KH2 PO4 , 0.2; (NH4 )2 SO4 , 0.14; urea, Semi-synthetic (LF) [200]
0.03; MgSO4 ·H2 O, 0.03; CaCl2 , 0.03; peptone,
0.1; crude cell wall of fungi, 1.0; plus trace metal
solution (1.0 ml)
Organic substrate medium Farm yard manure, rice chaffy grains, dried Complex (SSF) organic substrate [178]
banana leaf, banana pseudostem and rice bran
(500 g) and 100 ml of 30% molasses solution (v/v)
This table contains different media used for Trichoderma spp. BCA production.
a Valid for all media types.
al. [33,137] have reported production of T. viride on munici- mately 1–1000 kg/ha of dewatered sludge depending upon the
pal wastewater from BCA point of view. The studies advocated selected raw material. Thus, Trichoderma spp. production on
potential utilization of municipal wastewater sludge and indus- sludge would not just serve as potent BCA, but also as a novel
trial wastewaters as shown in the mass balance flow chart technique for sustainable sludge management.
(Fig. 3) according to the proposed estimation of use of sludge. These wastes often contain certain pollutants like, pesticides,
It was assumed for the schematic comparison between conven- metals complexes, whilst Trichoderma spp. are quite efficient
tional and alternative routes that Trichoderma spp. fermented in degrading these pollutants (e.g., organochlorine pesticides)
broths of conventional media/wastewaters/wastewater sludges [139]. They can tolerate high metal content by chelating metal
will be mixed with either dry Talc/silica powder, or dry dewa- ions [62]. The aforestated fact strengthens and encourages the
tered sludge powder as carrier material in the ratio of 1:99 ideology of using these wastes as substrates for Trichoderma
(w/w), respectively. Eventually, for conventional route (con- spp. based BCAs. Furthermore, wastewater and wastewater
ventional medium—108 CFU/ml) either extensive drying would sludges contain various complex compounds along with the
be required or the process may not be viable to obtain coni- essential components, hence there are ample opportunities that
dia concentrations ≥107 CFU/g formulated powder. On the one could encounter compound(s) that support(s)/induce(s)
other hand, alternative route (wastewater sludges/wastewater sporulation, in addition to vegetative growth of the fungi.
sludges—107 to 1010 CFU/ml) would require moderate air dry- Above all, large requirements of Trichoderma spp. based
ing. Therefore, for field application of fungal agents at a rate BCAs (50% of fungal BCAs [4]) and their subsequent demands
of 1013 CFU/ha [138] would warrant consumption of approxi- for enormous amounts of wastes will serve two purposes: (1)
Fig. 3. Schematic comparison of conidia requirements in conventional and alternative (non-conventional) routes of Trichoderma production.
sustainable Trichoderma spp. based BCAs production and aged and emphasized, thereby, facilitating standardization and
(2) waste management in a novel and environment-friendly commercialization of Trichoderma spp. based BCA. In fact,
way (reduced green house gases via incineration; and reduced bioassay methods need to be made more simpler, quantitative
hazard during land spreading-Trichoderma treated). and standardized for better comparisons.
5.5. Standard bioassay for quality control 5.6. Trichoderma spp. based formulations
Last but not the least, similar to other biopesticides, standard For obvious reasons, formulation development of BCAs is
bioassay technique for Trichoderma spp. is mandatory. How- one of the most important steps in the overall production process.
ever, this arena of Trichoderma spp. based BCAs is perhaps least Ideally, formulation ensures protection of the active ingredients
considered by researchers. Bioassay protocols simulating in situ (spores, conidia, mycelial germlings of antagonistic fungi) from
environment under in vitro conditions by best possible means extreme pHs, lower humidity, chemicals and UV radiation. For-
(diet incorporation technique, droplet test) for Trichoderma, as mulated BCAs exhibit antagonistic action without being affected
standard is still awaited. Meanwhile, literature is replete with by the adverse environmental factors. Many researchers have
either, rudimentary protocols, such as, measurement of inhi- reported different types of formulations of Trichoderma, e.g.,
bition zone and viable spore count [15,67,80–82,140], lytic invert emulsion [146]; cane molasses amendment [147]; seed
enzyme activity/metabolites [10,11] or, cumbersome green- coating [148]; pregelatinized starch-flour granules [149]; wet-
house/field study [82,141]. Consequently, there are several table powder [64]; alginate pellets [108]; gluten matrix [150];
examples where in vitro high potential Trichoderma spp. failed and application techniques, e.g., seed treatments [151,152]; soil
under in situ. Fortunately, there are occasional reports on amendments [153]; honey bee [154]; bumble bee [154]; com-
development of simpler bioassay techniques for Trichoderma posted chicken manure [16]; composted cow manure [155].
spp., which could simulate precise in situ conditions, such as, Lewis et al. [156] reported that T. hamatum and T. virens were
microdilution method based on infinite inhibition concentration effective in disease prevention (>80%) and pathogen reduc-
[142–144]; and tubular bioassay system to measure production tion (>75%) under greenhouse studies when amended with bran
of antagonistic metabolites at the antagonist–pathogen interface flakes or alginate pellets. However, in general, conidia without
[83]. Furthermore, for initial screening of potential Trichoderma any amendments were ineffective. Furthermore, Bae and Knud-
spp. for biocontrol, a reliable, image analysis based quantitative sen [108] and McLean et al. [157] have demonstrated that when
evaluation bioassay of in vitro antagonism was also reported introduced in soil, the biocontrol activity of Trichoderma spp.
[145]. However, further bioassay research needs to be encour- was formulation dependent.
Table 5
Comparison between Bacteria and Trichoderma spp. based BCAs
Bacteria Trichoderma sp.
Mode of action on pests through ingestion [136] Usually, mode of action on pests/weeds through contact [104]
Mass production normally by liquid fermentation [136] Mass production normally by solid-state fermentation [118]
They have been used extensively in field [187] Limited reported uses and market exploitation [4]
Many have been reported as plant growth enhancers, e.g., Rhizobium [201] Promote general plant growth and nutrition too [104]
Limited use in bioremediation of recalcitrants [202] Widely exploited in bioremediation methods of recalcitrants [28]
5.7. Integrated pest management with Trichoderma spp. neous biocontrol and growth promotion) of Trichoderma spp.
based BCAs are driving factors for their steadily cumulating
Many researchers have compared the efficiency of success.
Trichoderma based BCAs with conventional chemical fungi-
cides [64,152,155,158,159]. In comparison to chemical
7. Conclusions
fungicide—azoxystrobin, the biocontrol showed by T. virens
was as low as 33.3% disease infestation with 0.3% severity
Trichoderma spp. play major role as biocontrol agents, owing
according to Fisher’s LSD test (P = 0.05) [64]. Using fac-
to their capabilities of ameliorating crop-yields by multiple role,
torial design for in vitro experiment and randomized block
such as biopesticide, bioherbicides and plant growth promotion.
design for field experiment, integration of T. harzianum
Information on the classification of the genus, Trichoderma,
(105 spores/ml/g seed) and carboxin (2 g/kg seed) for seed
mechanisms of antagonism and role in plant growth promotion
treatment resulted in enhanced seed germination (12.0–14.0%)
has been well documented. However, fast paced current research
and grain yields (42.6–72.9%) and reduced wilt incidence
in this field should be carefully updated for the fool-proof com-
(44.1–60.3%) [158]. Similarly, other researchers have reported
mercialization of the fungi. In order to enhance marketability of
that the biocontrol exhibited by many Trichoderma spp. was
these fungi as BCAs, feasible commercial production processes
greater than chemical fungicides. Thus, it could be con-
are of utmost importance. Pursuit for cheaper and alternative
cluded that Trichoderma based BCAs were competitive to
substrates and optimal operating parameters to increase coni-
their conventional chemical counterparts. Meanwhile, combi-
dia production is on, and several encouraging results are being
nation of Trichoderma based BCAs and chemical fungicides
reported by researchers worldwide. Thus, it is expected that in
(integrated control) have been reported to be much more effec-
near future, exploitation of these interesting BCAs would be
tive than either of the two alone. In fact, integrated control
maximized.
by means of Trichoderma and chemical pesticides combi-
nations is becoming popular owing to cost economics as
well as sustainable approach [152,158,159]. Furthermore, use Acknowledgements
of Trichoderma in integrated pest management makes them
potential partners in pest control for quick and safer mea- The authors are sincerely thankful to Natural Sciences
sures. and Engineering Research Council of Canada (Grants A4984,
STP235071, Canada Research Chair) for financial support.
6. Market potential The views and opinions expressed in this article are those of
authors. Last but not the least, the authors would also like
Presently, Trichoderma spp. based products are considered as to thank Dr. Guillemond Ouellette (Histology, Host-pathogen
relatively novel type of BCAs. In comparison to B. thuringien- Interaction—Natural Resources Canada) for his valuable sug-
sis (Bt) biopesticides, their market size is quite small (Bt shares gestions to this work.
about 97% of overall biopesticides), they fall in remaining 3%
bracket, which also comprises viral and nematode based biopes- References
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Biochemical Engineering Journal 37 (2007) 1–20

Antagonistic fungi, Trichoderma spp.: Panoply of biological control

1. Introduction trum in terms of disease control and production yield [5]. In

M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20

M. Verma et al. / Biochemical Engineering Journal 37 (2007) 1–20

strawberries [106]. Despite all the research, there is a need for

Co-metabolism in the presence of glycerol

Co-metabolism in the presence of glucose

Mycorrhizal fixation of phosphorus and

directly uptaking organic N from soil

well defined or, no microflora present) tomato, thereby, showing

growth promotional ability of this fungus. The physical pres-

based BCAs for growth promotion are enhanced due to symbio-

easily acquire nutrients from complex substrates like, protein-

tannin, and glucosamine, in soil due to their ectomycorrhizal

Rhizosphere enhancers—N and P

land, consequently affecting plant yield. The positive role of

orated by Wu et al. [110] which is an indirect mode for their

plant growth promotional activity.

3.2. Indirect effect

100 ␮g/g soil

3.2.1. Induction of plant defence mechanism

choderma with roots, and several lytic enzymes induced by plant

Strawberry, Fragaria ananassa

defence system destroyed and consumed the pathogen cell wall.

Finally, it provided nutrient to the plant as depicted in Fig. 1.

A mycoparasitic interaction study in this context revealed that

ity of different mechanisms of induction for each type of lytic

enzyme and the necessity to characterize these enzymes and

T. longibrachiatum, T. harzianum, T. viride,

BCA assessment is difficult due to multiple factors associated

and scarce information available. Nevertheless, there are many

Trichoderma spp. based commercial products in market which

The concept of utilizing fungi for bioremediation of soil

is ample evidence of various Trichoderma spp. contribut-

ing to polycyclic aromatic hydrocarbons (PAHs) degradation,

whilst affecting native mycorrhizal fungi both positively and/or,

U.S., U.S., Belgium

negatively [112]. Saraswathy and Hallberg [113] reported a max-

Antagonism by enzymes and antibiotics New Zealand

Katayama and Matsumura [28] demonstrated degradation

Mycoparasite living on other fungi

and dichlorodiphenyl trichloroethane (DDT). Thus, Tricho-

other lytic enzymes produced in abundance by Trichoderma

Rhizoctonia solani, Sclerotium rolfsii, Pythium

to be curtailed. However, this can be achieved, if and only if the

Armillaria, Botryosphaeria and other fungal

contaminated soil is inoculated with Trichoderma spp. at regu-

lar defined intervals. In fact, inclusion of Trichoderma spp. in

“integrated pest management” program is a must. In the current

on nursery and field crops

needs of agricultural commodities.

Therefore, consistent simultaneous application of some

ful pesticides/herbicides; the former can tolerate and degrade

would provide an agreeable soil environment. This will assure

J.H. Biotech, Mycontrol, Ltd., De

on consistent-simultaneous use of Trichoderma spp. as BCA

5.1. Microbial propagules

Source: http://www.google.com internet search engine.