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.

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