TERM PAPER ON

TRICHODERMA VIRIDE

SUBMITTED BY: SHASHI SHARMA

SECTION: P8003
ROLL NO. : P8003B15
REGISTRATION NUMBER: 11006142

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 CONTENTS

1. INTRODUCTION

2 OBJECTIVE TO STUDY

3 INDUSTRIAL IMPORTANCE

4FUTURE PROSPECTS

5 REFRENCES

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INTRODUCTION: TRICHODERMA VIRIDE is a genus of fungi that is present in all soils
where they are the symbionts .many species in this genus can be characterized as
opportunistic a virulent plant.There are 89 species of Trichoderma genus .some of these are
as following and everyone has its own importance.

 Trichoderma aggressivum
 Trichoderma asperellum
 Trichoderma atroviride
 Trichoderma aureoviride
 Trichoderma austrokoningii
 Trichoderma brevicompactum
 Trichoderma candidum
 Trichoderma caribbaeum var. aequatoriale
 Trichoderma caribbaeum var. caribbaeum
 Trichoderma catoptron

Cultures are typically fast growing at 25-30°C, but will not grow at 35°C. Colonies are
transparent at first on media such as cornmeal dextrose agar (CMD) or white on richer media
such as potato dextrose agar (PDA). Mycelium are not typically obvious on CMD, conidia
typically form within one week in compact or loose tufts in shades of green or yellow or less
frequently white. A yellow pigment may be secreted into the agar, especially on PDA. Some
species produce a characteristic sweet or 'coconut' odor.

Conidiophores are highly branched and thus difficult to define or measure, loosely or
compactly tufted, often formed in distinct concentric rings or borne along the scant aerial
hyphae. Main branches of the conidiophores produce lateral side branches that may be paired

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or not, the longest branches distant from the tip and often phialides arising directly from the
main axis near the tip. The branches may rebranch, with the secondary branches often paired
and longest secondary branches being closest to the main axis. All primary and secondary
branches arise at or near 90° with respect to the main axis. The
typical Trichoderma conidiophore, with paired branches assumes a pyramidal aspect.
Typically the conidiophore terminates in one or a few phialides. In some species (e.g. T.
polysporum) the main branches are terminated by long, simple or branched, hooked, straight
or sinuous. thin-walled, sterile or terminally fertile elongations. The main axis may be the
same width as the base of the phialide or it may be much wider.: A total of 26 morphological
and colour mutants ofTrichoderma viride were characterized. They were divided into three
groups based on morphology, pigmentation, growth rates and intensity of conidiation.
Complementation analysis of colour mutants and mutants with disturbances in conidiation
showed that after anastomosis and protoplast fusion only heterokaryotes are formed while no
nuclear migration and diploidization takes place

CONIDIOSPORES OF TRICHODERMA VIRIDE;

TRICHODERMA VIRIDE is found all above in nature.Mostly species are isolated from
forest or agricultural soil at all latitudes. HYPOCREA species are most frequently found on
bark or decorticated of wood but many species grown on bracket fungi.ex:H .pulvinate
H.avellanea.

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OBJECTIVE TO STUDY:

Fungi of the genus Trichoderma are soilborne, green-spored ascomycetes that can be found

all over the world. They have been studied with respect to various of their habitats, efficiently
fighting their competitors. Once established, they launch their potent degradative machinery
for decomposition of the often heterogeneous substrate at hand. Therefore, distribution and
phylogeny, defense mechanisms, beneficial as well as deleterious interaction with hosts,
enzyme production and secretion, sexual development, and response to environmental
conditions such as nutrients and light have been studied in great detail with many species of
this genus, thus rendering Trichoderma one of the best studied fungi with the genome of three
species currently available. Efficient biocontrol strains of the genus are being developed as
promising biological fungicides, and their weaponry for this function also includes secondary
metabolites with potential applications as novel antibiotics. The cellulases produced
by Trichoderma reesei, the biotechnological workhorse of the genus, are important industrial
products, especially with respect to production of second generation biofuels from cellulosic
waste. Genetic engineering not only led to significant improvements in industrial processes
but also to intriguing insights into the biology of these fungi and is now complemented by the
availability of a sexual cycle inT. reesei/Hypocrea jecorina, which significantly facilitates
both industrial and basic research. This review aims to give a broad overview on the qualities
and versatility of the best studied Trichoderma species and to highlight intriguing findings as
well as promising applications.
Trichoderma spp. are highly successful colonizers of their habitats, which is reflected both by
their efficient utilization of the substrate at hand as well as their secretion capacity for
antibiotic metabolites and enzymes. They are able to deal with such different environments as
the rich and diversified habitat of a tropical rain forest as well as with the dark and sterile
setting of a biotechnological fermentor or shake flask. Under all these conditions, they
respond to their environment by regulation of growth, conidiation, enzyme production, and
hence adjust their lifestyle to current conditions, which can be exploited for the benefit of
mankind. One of these environmental factors is the presence or absence of
light. Trichoderma has a long tradition of research toward the effect of light on its physiology
and development, which already started in 1957 and largely paralleled that of Phycomyces
blakesleeanus .

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In recent years, research with Trichoderma has been facilitated significantly by sequencing of
the genomes of three strains representing the most important applications of this genus: The
genome sequence of T. reesei, the industrial workhorse. surprisingly revealed that, despite its
importance in industrial cellulase production, its genome comprises the fewest amount of
genes encoding cellulolytic and hemicellulolytic enzymes. Analysis and annotation of the
genomes of Trichoderma atroviride and Trichoderma virens, two important biocontrol
species is still in progress. Interestingly, the genomes of T. atroviride and T. virens are
significantly larger than that of T. reesei, and they comprise roughly 2000 genes more than
does T. reesei. It will be interesting to learn the significance of this considerable difference in
genome sizes in the physiology of these fungi. These milestones in research
with Trichoderma enabled detailed studies, which provided intriguing insights into their
lifestyle, physiology, and the underlying mechanisms at the molecular level .

INDUSTRIAL IMPORTANCE OF TRICHODERMA VIRIDE

Trichoderma, being a saprophyte adapted to thrive in diverse situations, produces a wide

array of enzymes. By selecting strains that produce a particular kind of enzyme, and culturing
these in suspension, industrial quantities of enzyme can be produced.

1. T. reesei is used to produce cellulase and hemicellulase 

2. T. longibratum is used to produce xylanase 
3. T. harzianum is used to produce chitinase.

Biocontrol agent

Several strains of Trichoderma have been developed as biocontrol agents against fungal

diseases of plant.] The various mechanisms include antibiosis, parasitism, inducing host-plant
resistance, and competition. Most biocontrol agents are from the species T. harzianum, T.

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viride and T. hamatum. The biocontrol agent generally grows in its natural habitat on the root
surface, and so affects root disease in particular, but can also be effective against foliar
disease
USES IN THE PRODUCTION OF ALPHA-AMYLASE:

The production of extracellular α-amylase by the mesophilic fungus Trichoderma viride was

optimized in shake-flask cultivation. Overall, the data imply that α-amylase from fungus was
induced by a variety of starchy substrate but maximum enzyme production was stimulated by
corn starch at a concentration of 4.0%. Enzyme synthesis was affected by nitrogen sources,
and maximal activity was attained with inorganic than organic nitrogen sources. Effect of
different salts in the culture medium was evaluated and it was found that Mg2+, Fe2+, K+ and,
to some extent, Ca2+ and Na+ play an important role for optimum production of α-amylase by
the fungus. Maximum enzyme production was obtained after 3 days of incubation in a
fermentation medium with initial pH 5.0 at 30° under continuous agitation at 180 rpm. These
propertiesake the enzyme suitable for industrial uses.

USED IN STARCH INDUSTRRY:

Starch industry wastewater was investigated to assess and improve its potential as a raw
material for the conidia production of biocontrol fungi, Trichoderma viride. The wastewater
was tested with and without supplements of glucose, soluble starch, meat peptone and
probable conidiation inducer chemicals in shake flask culture. Addition of complex carbon
source (soluble starch, 1% and 2% w/v) produced maximum conidia (≈3.02 and
4.2 × 1010 CFU/mL, respectively). On the other hand, glucose addition as a simpler carbon
source was either ineffective or, reduced conidia production (from 1.6 × 108 in control to
3.0 × 107 CFU/mL in 5% w/v glucose supplement). Supplement of nitrogen source showed a
small increase of conidia concentration. Propionic, maleic and humic acids, EDTA, pyridine,
glycerol and CaCO3 were examined as probable conidiation inducers and showed effect only
on initial rate of conidiation with no increase in final conidia concentration. Intra and
extracellular ATP correlation with spore production showed dependence on growth media
used and conidia concentration at the end of fermentation. Addition of carbon and nitrogen
sources showed an increase in protease activity (from 0.4985 to 2.43 IU/mL) and
entomotoxicity (from 10448 to 12335 spruce budworm unit (SBU)/μL). Entomotoxicity was
improved by 11% in fermenter over shake flask when starch industry wastewater was
supplemented with meat peptone.

AS A BIOPESTICIDES:

In the recent years, the environmental contamination caused by excessive use of chemical
pesticides increased the interest in integrated pest management, where chemical pesticides are

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substituted by biopesticides to control plant pests and plant diseases. Trichoderma-Based
Biocontrol Agents (BCAs) possess better ability to promote plant growth and soil
remediation activity compared to their counterparts (virus, bacteria, nematodes and protozoa
(3), (4). Their capability to synthesize antagonistic compounds (proteins, enzymes and
antibiotics) and micro-nutrients (vitamins, hormones and minerals) enhance their biocontrol
activity.
Like other fungal BCAs, conidial mass of Trichoderma is the most proficient propagule,
which tolerates downstream processing (e.g., air drying). Despite the advantages, mass
production of Trichoderma BCAs is less prevalent, owing to high-cost raw materials like
Mendel's medium, molasses, corn steep liquor and other.

Shortly after the discovery of T. viride QM6a by the US army during World War II
T.Reese the outstanding efficiency of its cellulases led to extensive research toward industrial
applications of these enzymes. Later on, this species was renamedT. reesei in honours of
Elwyn species T. Reese) and became the most important cellulase producer worldwide.

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USED AS CELLULASES AND PLANT CELL WALL DEGRADING ENZYMES:

Rising energy costs and the imminent climate change led to an increased attention to
biofuel production . As a potent cellulase producer, research with T. reesei is nowadays
particularly focused on improvement of efficiency of the enzyme cocktail produced in order
to decrease overall costs of production of bioethanol from cellulosic waste material , although
applications in the pulp and paper industry and textile industry are also important. After the
early mutation programs and strain improvement, the protein secretion capacity of industrial
strains now reaches 100 g/l, with up to 60% of the major cellulase Cel7a (CBHI) and 20% of
Cel6a (CBHII). High levels of cellulase and hemicellulase gene expression can be achieved
upon cultivation on cellulose, xylan, or a mixture of plant polymers as well as on lactose all
of which are agricultural or industrial byproducts. The natural inducer of at least a subset of
these enzymes is believed (yet not definitely proven) to be sophorose, a transglycosylation
product of cellobiose . Targeted strategies to further enhance the efficiency of the enzymes
secreted include elucidation of regulatory mechanisms both at the promotor level as well as
with respect to signal transduction .However, auxiliary components acting on the substrate
could also enhance efficiency of its degradation .

FUTURE PROSPECTS:

More than ever before sustainable economy and protection of our environment are dominant
topics in our everyday life and one alarming report about contaminated landscapes or
catastrophes caused by climate change follows another. Today, 87% of energy used in the
world comes from nonrenewable sources like natural gas, oil, and coal. Although biofuel
production is now being pushed in order to decrease the requirement for fossil fuels, the raw
materials therefore originate from commodities and land also needed for food. In this respect,
production of the so-called second generation biofuels from agricultural waste products by
the aid of cellulases and hemicellulases produced for example by T. reesei and fermentation
of the resulting oligosaccharides by yeast provides an alternative strategy. However, for an
economically competitive process an increase in efficiency of more than 40-fold would be
necessary, which is a formidable challenge for research with Trichoderma.

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Sustainability is also the major driving force for investigation of biocontrol withTrichoderma.
As opportunistic plant symbionts and effective mycoparasites, numerous species of this genus
have the potential to become commercial biofungicides. The challenge in this field of
research will be the development of reliable screening techniques, which allow for prediction
of the biocontrol efficiency of a given isolate by determination of the key factors for this
process. Nevertheless, also the ecological effects of widespread application of a single (or
few) fungal species in agriculture remain to be investigated in order to ensure a truly
beneficial effect for the environment.

Besides these major applications of Trichoderma spp., also the fields of green and white
biotechnology become increasingly important for environmentally safe production of
enzymes and antibiotics. These industrial applications will also benefit from studies on
molecular physiology and regulatory processes, which continuously reveal novel and
valuable metabolites and enzymes as well as components to be modified or adjusted for cost
effective high yield production.

Last but not least, the extensive studies on diverse physiological traits available and still
progressing for Trichoderma make these fungi versatile model organisms for research on
both indudustrial importance.

 REFRENCES:

 Adrio JL, Demain AL. Fungal biotechnology. Int Microbiol.  2003;6:191–199. 

 Alanio A, Brethon B, Feuilhade de Kerviler Chauvin M, E, Leblanc T, Lacroix C,

Baruchel A, Menotti J. Invasive pulmonary infection due to Trichoderma
longibrachiatum mimicking invasive Aspergillosis in a neutropenic patient
successfully treated with voriconazole combined with caspofungin. Clin Infect
Dis. 2008;46:e116–e118. 

 Antal Z, Kredics L, Pakarinen J, Doczi I, Andersson M, Salkinoja-Salonen M,

Manczinger L, Szekeres A, Hatvani L, Vagvolgyi C, Nagy E. Comparative study of
potential virulence factors in human pathogenic and saprophytic Trichoderma
longibrachiatum strains. Acta Microbiol Immunol Hung. 2005;52:341–350.

 Antal Z, Varga J, Kredics L, Szekeres A, Hatvani L, Manczinger L, Vagvolgyi C,

Nagy E. Intraspecific mitochondrial DNA polymorphism within the emerging
filamentous fungal pathogen Trichoderma longibrachiatum. J Med
Microbiol.  2006;55:31.

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  Baek JM, Kenerley CM. The arg2 gene of Trichoderma virens: cloning and
development of a homologous transformation system. Fungal Genet
Biol.  1998;23:34–44. 

 Bansal P, Hall M, Realff MJ, Lee JH, Bommarius AS. Modeling cellulase kinetics on
lignocellulosic substrates. Biotechnol Adv. 2009;27:833–848. 

 Benitez T, Rincon AM, Limon MC, Codon AC. Biocontrol mechanisms

of Trichoderma strains.Int Microbiol. 2004;7:249–260. 

 Blumenthal CZ. Production of toxic metabolites in Aspergillus niger, Aspergillus

oryzae, andTrichoderma reesei: justification of mycotoxin testing in food grade
enzyme preparations derived from the three fungi. Regul Toxicol
Pharmacol. 2004;39:214–228. 

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