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““TRICHODRMA AS A BIOCONTROLTRICHODRMA AS A BIOCONTROL
AGENTAGENT ""
Avs trichodrma  as a biocontrol  agent
 What is biological control, what are the benefits
to its use
 Need of biological control agents
 Mechanism of biological control agents
 Requirements of successful biocontrol agents
 Working example of biocontrol
agentTrichoderma ssp.
 Available Biopesticides and its use in
Agriculture
Importance of plant diseases
Estimated annual crop production worldwide
Amount lost to disease, insects, weeds using
current control measures
Additional losses without current control measures
$1.2 - 1.3 trillion$1.2 - 1.3 trillion
$500 billion$500 billion
$330 billion$330 billion
Verticillium wiltVerticillium wilt
Citrus cankerCitrus canker
Fireblight of pearFireblight of pear Apple scab
Rice blast
( IMF and CIA World Factbook, 2014)
2013: world spent $43.4 billion$43.4 billion on
chemical pesticides
Of this, < 1%< 1% actually gets to
where the pathogen is
What happens to the rest?
Ground water
Taken up by the plant
Development of resistance
     Current approaches to disease control
Chemical
Identification of resistance genes
Introgressing into commercial
cultivars
Problems with development of
resistance, pyramiding genes
Breeding
Biological Control is an attractive alternative/supplement
Control of plant pathogens and diseases caused by them through antagonistic
microorganisms or botanicals is termed biological control agents
 According to Baker and Cook’s (1974) - “Biological control is the
reduction of inoculum or disease producing activity of a pathogen
accomplished by or through one or more organisms other than man.”
 Antagonistic microorganisms like species of Trichoderma, Penicillium,
Bacillus, Pseudomonas etc.
 Chemical pesticides
Implicated in environmental and human health
problems
– Require yearly treatments and expensive
– Toxic to both beneficial and pathogenic species
• Biological control agents
– Non-toxic to human and animal
– Not polluted
– Host specific
•Only effect one or few species
WHEN :
 Biological control agents are
◦ Low cost
◦ Labor intensive
◦ Host specific
WHILE :
 Chemical pesticides are:
◦ cost-effective
◦ easy to apply
◦ Broad spectrum
How does Biological Control works
Nutrient Competition
Biological
Control
Antibiosismycoparasitism
Induced resistance
Tolerance to stress
through enhanced
root and plant
development
Inactivation of the
pathogen’s enzymes
 An ideal biocontrol agent should satisfy
most all, of the following attributes
• Must not be pathogenic to plants and animals
• Level of pathogen control must be high
• Should live longer in soil or host tissues
• Should have rapid reproductive capacity
• Should be a good competitor
• Should have high survival rate in soil or host tissues
• Should be capable of controlling more than one
pathogen
• Should be suitable for long-term storage
• Should be compatible to use with agro-chemicals
viz. fertilizers, pesticides etc.
Avs trichodrma  as a biocontrol  agent
1671 – First found in Germany
1794 – Identified by Persoon almost 218 years ago
1927 – Gilman and Abbott recognized four species based on
colour, shape of conidia and colony appearance
>75 years ago the potential use of Trichoderma by Weindling
(1932) and first to demonstrate the parasitic activity in wilt of
Pigeon pea
Best known mycoparasite against many soil borne plant pathogens
 Very effective biological agent
 Free living
 Highly proliferating
 Non- pollutive
 Easily accessible
 Non phytotoxic
 Systemic ephemeral
 Readily biodegradable
 Cost effective
 Synergistic effect
 Longer shelf life
 Greater compatibility
 A genus of fungi, including many species that can be used
to control phytopathogenic fungi.
 generally, soil dwelling saprophytes. They have a rapid
growth rate, sporulate abundantly, compete well with other
 show resistance to chemical pesticides and produce various
antibiotics (e.g., gliotoxin and viridin).
 control of wood-rotting, wound-infecting and soil-borne
fungal pathogens of seedlings and mature plants.
Conidiophores
Conidia
Hyphae
Morphological structure of Trichoderma
Kulkarni and Sagar (2007) mentioned the Trichoderma as asexual stage
and Hypocrea as sexual stage
Position Asexual stage
(conidia)
Sexual stage
(ascospore)
Kingdom Fungi Fungi
Phylum Ascomycota Ascomycota
Sub-division Deuteromycotina Ascomycotina
Class Hyphomycetes Pyrenomycetes
Order Monilliales Sphariales
Family Monilliaceae Hypocreaceae
Genus Trichoderma Hypocrea
 Cultures are fast growing at 25-30° C
 Conidia forming within on week in compact
or loose tufts in shades of green or yellow
or less frequently white
 Yellow pigment may be secreted into the
agar, specially on PDA
 A characteristic sweet or ‘coconut’ odour
is produced by some species
Chlamydospores
Conidia
Phialides
Hyphae
Conidiophores
Septate hyaline hyphae.
Conidiophores are
hyaline, branched
Phialides are hyaline,
flask-shaped, and inflated
at the base.
The colour of
the conidia is mostly
green.
Trichoderma spp.
may also produce
chlamydospores
Singh et al., 2007.
Potential bio control activities exhibited by TrichodermaPotential bio control activities exhibited by Trichoderma
Kamala and Indira, 2012, Manipur
For space and nutrients under specific condition do not get substrate
Suppress growth of pathogen population
e.g: Soil treatment with Trichoderma harzianum spore suppressed
infestation of Fusarium oxysporum f. sp. vasinfectum and F. oxysporum f.
sp. melonis
(Perveen and Bokhari, 2012)
Mechanisms of action
Mycoparasitism
Antagonist fungi parasitize other pathogenic fungi
Hyphae of Trichoderma either grow along the host hyphae or coil
around it
E.g. : T. harzianum and T. hamatum were mycoparasite
of both Scelerotium rolfsii and R. solani
Interaction –
 Coiling of hyphae around the pathogen,
 Vacuolization,
 Penetration by haustoria and
 lysis (Omero et al., 1999).
 Recognize and attach to the pathogenic fungus and excrete extra-cellular
lytic enzymes like β-1,3-glucanase, chitinase, proteases and lipase
(Schlick et al., 1994).
Trichoderma coils around, penetrates, and kills other fungi that are
pathogenic (i.e. cause disease) to crops. It can digest their cell walls
A clear view with an electron microscope
Trichoderma spp.(T) fungal strands
coil (C) around the Rhizoctonia (R)
Initial stages of degradation (D) as a result of
Trichoderma generated enzymes.
T: Trichoderma R: Rhizoctonia
Antibiosis
 It is the condition in which one or more metabolites
excreted by an organism have harmful effect on one or more
other organisms
 In such antagonistic relationship spp. A produces a chemical
substance that is harmful to Spp. B without a Spp. A deriving
any direct benefit
e.g: Trichoderma secreted - Trichodermin, viridine,
Trichothecin, Sesqiterpine etc.
Growth inhibition of R. solani by the T. virens produced antibiotic
gliotoxin . A: Gliotoxin amended B: non amended
Cont…
Trichoderma strains solubilize phosphates and micronutrients
The application of Trichoderma strains in rhizosphere of plants
increases the number of deep roots, there by increasing the plants
ability to resist drought
Plant growth promoter
Fig.: Enhanced root development from field grown bean plants as a consequence
of root colonization by the rhizosphere competent strain T. harzianum
(Amin et al., 2010)
Cont…
Avs trichodrma  as a biocontrol  agent
Mass production of biocontrol agents
Liquid fermentation method
Avs trichodrma  as a biocontrol  agent
 Substrates for mass multiplication: wheat bran, wheat
straw, FYM, press mud, coir pith, ground nut shell, rice bran, etc
 Carrier/ food base materials: Talc, vermiculite, molasses,
gypsum, kaolin, peat, sodium alginate, Cacl2
Organism Trade Name Target Crops
Trichoderma
virens
SoilGard 12G3 Pythium,
Rhizoctonia, and
Root rots
Ornamental and food crop plants grown in
greenhouses,
Trichoderma
harzianum Rifai
strain KRL-AG3
Plant Shield® HC Fusarium,
Pythium, and
Rhizoctonia
Cucurbit vegetables, flowers, bedding plants,
Trichoderma
harzianum Rifai
strain KRL-AG2
T-22™ HC Fusarium,
Pythium, and
Rhizoctonia
Agronomic field and row crops, alfalfa, hay and
forage crops, bulb crops, cucurbits, fruiting
vegetables, herbs, spices, leafy vegetables,
cole crops, legumes, root crops, small grains
and tuber crops
Trichoderma
harzianum Rifai
strain KRL-AG2
T-22™ Planter
Box
Fusarium,
Pythium, and
Rhizoctonia
Agronomic field and row crops, alfalfa, hay and
forage crops, bulb crops, cucurbits, fruiting
vegetables
 There are several reputable companies that manufacture
government registered products.
Trade Name Bio agent Manufacture
Eco fit T. viride Hoechst and Schering AgroEvo Ltd,
Mumbai India
Super visit T. harzianum Fytovita, Czech Republic
Soil guard T. virens Certis Inc,Columbia,MD,USA
Root pro T. harzianum Efal Agri, Netanyl,Israel
Tusal T. Viride +T.
harzianum
Tusal Carrera Ester, Lleida Spain
Agroderma, Bio-cure,
Bioderma, Ecofit,
Rakshak, Trichosan
Trichoderma viride
Biocure (B&F) T. Viride and P. flourescens
1.Seed treatment
•Dose: @5 g /kg of seed
•Method: Make a paste or slurry adding 5 g in 10-20 ml of water
. Pour 1kg of seed on to the paste or slurry and mixed properly
to coat the seeds uniformly .Shade dry the coated seeds for 20-
30 minutes before sowing
2.Tuber/Rhizome/Cutting treatment:
•Dose: @ 10 g /litre of water
•Method:Dip the tuber / rhizome/cuttings in the suspension
prepare @ 10 g /litre of water. Shade dry for 15 minutes before
planting
(Medhi,2009)
Seedling Treatment:
•Dose: 300 g/ha.
•Method: Prepare a suspension @ 5-10 g/litre of water. Dip the
roots of seedling for 15minutes and shade dry for 15 minutes
before transplanting
Nursery bed treatment:
•Dose: 250 g for 400 sq.m nursery bed.
•Method: 1) prepare a suspension by adding 250 g in 50 litres of
water and drench the nursery bed soil.
•2) Mixed 250 g in 2 kg cow dung / compost/FYM and spread
over 400 sq.m. nursery bed and irrigate the bed.
Soil treatment:
i) Direct broadcasting:
•Dose: 300 g/ha.
•Method: Mix 300 g in 6 kg of FYM. Broadcast in one ha of land and irrigate the
field.
ii) Awaited broadcasting
•Dose: 30 g/ha
•Method: Mixed 30 g in 6 kg of FYM. Cover the mixture with polythene sheet for 7-
15 days and broadcast in the field
iii) Furrow application:
•Dose:300g /ha.
•Method: It is highly effective in root crops like potato, ginger, turmeric etc and
sugarcane.The mixture is applied in furrows at the time of earthing up or after 30
days of planting
• Decrease disease intensity.
• Reduce the use of chemical fungicides.
•Reduce undesirable effects from chemical pesticide.
• Play a key role in integrated management of diseases
•Safe for the users and the farming community.
• Provide natural long term immunity to crops and
soil
( Shrivastava , 1996 ).
•Deterious effects on non-target micro-
organisms
•Pathogens may develop resistance to the
biocontrol agent
•Pathogen replacement may follow control of
target disease pathogen
•Seasonal/weather phenomena can make
biocontrol agent ineffective
( Shrivastava , 1996 ).
CONCLUSIONS
Biological control an attractive alternative to
chemicals
Trichoderma species are effective in biological control of
fungus-induced plant disease
Plant diseases cause major loss of food and
money
Biological control occurs via several mechanisms
Competition
Antagonism
Mycoparasitism
Avs trichodrma  as a biocontrol  agent

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Avs trichodrma as a biocontrol agent

  • 1. ““TRICHODRMA AS A BIOCONTROLTRICHODRMA AS A BIOCONTROL AGENTAGENT ""
  • 3.  What is biological control, what are the benefits to its use  Need of biological control agents  Mechanism of biological control agents  Requirements of successful biocontrol agents  Working example of biocontrol agentTrichoderma ssp.  Available Biopesticides and its use in Agriculture
  • 4. Importance of plant diseases Estimated annual crop production worldwide Amount lost to disease, insects, weeds using current control measures Additional losses without current control measures $1.2 - 1.3 trillion$1.2 - 1.3 trillion $500 billion$500 billion $330 billion$330 billion Verticillium wiltVerticillium wilt Citrus cankerCitrus canker Fireblight of pearFireblight of pear Apple scab Rice blast ( IMF and CIA World Factbook, 2014)
  • 5. 2013: world spent $43.4 billion$43.4 billion on chemical pesticides Of this, < 1%< 1% actually gets to where the pathogen is What happens to the rest? Ground water Taken up by the plant Development of resistance      Current approaches to disease control Chemical Identification of resistance genes Introgressing into commercial cultivars Problems with development of resistance, pyramiding genes Breeding Biological Control is an attractive alternative/supplement
  • 6. Control of plant pathogens and diseases caused by them through antagonistic microorganisms or botanicals is termed biological control agents  According to Baker and Cook’s (1974) - “Biological control is the reduction of inoculum or disease producing activity of a pathogen accomplished by or through one or more organisms other than man.”  Antagonistic microorganisms like species of Trichoderma, Penicillium, Bacillus, Pseudomonas etc.
  • 7.  Chemical pesticides Implicated in environmental and human health problems – Require yearly treatments and expensive – Toxic to both beneficial and pathogenic species • Biological control agents – Non-toxic to human and animal – Not polluted – Host specific •Only effect one or few species
  • 8. WHEN :  Biological control agents are ◦ Low cost ◦ Labor intensive ◦ Host specific WHILE :  Chemical pesticides are: ◦ cost-effective ◦ easy to apply ◦ Broad spectrum
  • 9. How does Biological Control works Nutrient Competition Biological Control Antibiosismycoparasitism Induced resistance Tolerance to stress through enhanced root and plant development Inactivation of the pathogen’s enzymes
  • 10.  An ideal biocontrol agent should satisfy most all, of the following attributes • Must not be pathogenic to plants and animals • Level of pathogen control must be high • Should live longer in soil or host tissues • Should have rapid reproductive capacity • Should be a good competitor • Should have high survival rate in soil or host tissues • Should be capable of controlling more than one pathogen • Should be suitable for long-term storage • Should be compatible to use with agro-chemicals viz. fertilizers, pesticides etc.
  • 12. 1671 – First found in Germany 1794 – Identified by Persoon almost 218 years ago 1927 – Gilman and Abbott recognized four species based on colour, shape of conidia and colony appearance >75 years ago the potential use of Trichoderma by Weindling (1932) and first to demonstrate the parasitic activity in wilt of Pigeon pea Best known mycoparasite against many soil borne plant pathogens
  • 13.  Very effective biological agent  Free living  Highly proliferating  Non- pollutive  Easily accessible  Non phytotoxic  Systemic ephemeral  Readily biodegradable  Cost effective  Synergistic effect  Longer shelf life  Greater compatibility
  • 14.  A genus of fungi, including many species that can be used to control phytopathogenic fungi.  generally, soil dwelling saprophytes. They have a rapid growth rate, sporulate abundantly, compete well with other  show resistance to chemical pesticides and produce various antibiotics (e.g., gliotoxin and viridin).  control of wood-rotting, wound-infecting and soil-borne fungal pathogens of seedlings and mature plants. Conidiophores Conidia Hyphae Morphological structure of Trichoderma
  • 15. Kulkarni and Sagar (2007) mentioned the Trichoderma as asexual stage and Hypocrea as sexual stage Position Asexual stage (conidia) Sexual stage (ascospore) Kingdom Fungi Fungi Phylum Ascomycota Ascomycota Sub-division Deuteromycotina Ascomycotina Class Hyphomycetes Pyrenomycetes Order Monilliales Sphariales Family Monilliaceae Hypocreaceae Genus Trichoderma Hypocrea
  • 16.  Cultures are fast growing at 25-30° C  Conidia forming within on week in compact or loose tufts in shades of green or yellow or less frequently white  Yellow pigment may be secreted into the agar, specially on PDA  A characteristic sweet or ‘coconut’ odour is produced by some species
  • 17. Chlamydospores Conidia Phialides Hyphae Conidiophores Septate hyaline hyphae. Conidiophores are hyaline, branched Phialides are hyaline, flask-shaped, and inflated at the base. The colour of the conidia is mostly green. Trichoderma spp. may also produce chlamydospores
  • 18. Singh et al., 2007.
  • 19. Potential bio control activities exhibited by TrichodermaPotential bio control activities exhibited by Trichoderma Kamala and Indira, 2012, Manipur
  • 20. For space and nutrients under specific condition do not get substrate Suppress growth of pathogen population e.g: Soil treatment with Trichoderma harzianum spore suppressed infestation of Fusarium oxysporum f. sp. vasinfectum and F. oxysporum f. sp. melonis (Perveen and Bokhari, 2012) Mechanisms of action
  • 21. Mycoparasitism Antagonist fungi parasitize other pathogenic fungi Hyphae of Trichoderma either grow along the host hyphae or coil around it E.g. : T. harzianum and T. hamatum were mycoparasite of both Scelerotium rolfsii and R. solani
  • 22. Interaction –  Coiling of hyphae around the pathogen,  Vacuolization,  Penetration by haustoria and  lysis (Omero et al., 1999).  Recognize and attach to the pathogenic fungus and excrete extra-cellular lytic enzymes like β-1,3-glucanase, chitinase, proteases and lipase (Schlick et al., 1994).
  • 23. Trichoderma coils around, penetrates, and kills other fungi that are pathogenic (i.e. cause disease) to crops. It can digest their cell walls A clear view with an electron microscope Trichoderma spp.(T) fungal strands coil (C) around the Rhizoctonia (R) Initial stages of degradation (D) as a result of Trichoderma generated enzymes. T: Trichoderma R: Rhizoctonia
  • 24. Antibiosis  It is the condition in which one or more metabolites excreted by an organism have harmful effect on one or more other organisms  In such antagonistic relationship spp. A produces a chemical substance that is harmful to Spp. B without a Spp. A deriving any direct benefit e.g: Trichoderma secreted - Trichodermin, viridine, Trichothecin, Sesqiterpine etc.
  • 25. Growth inhibition of R. solani by the T. virens produced antibiotic gliotoxin . A: Gliotoxin amended B: non amended Cont…
  • 26. Trichoderma strains solubilize phosphates and micronutrients The application of Trichoderma strains in rhizosphere of plants increases the number of deep roots, there by increasing the plants ability to resist drought Plant growth promoter
  • 27. Fig.: Enhanced root development from field grown bean plants as a consequence of root colonization by the rhizosphere competent strain T. harzianum (Amin et al., 2010) Cont…
  • 29. Mass production of biocontrol agents Liquid fermentation method
  • 31.  Substrates for mass multiplication: wheat bran, wheat straw, FYM, press mud, coir pith, ground nut shell, rice bran, etc  Carrier/ food base materials: Talc, vermiculite, molasses, gypsum, kaolin, peat, sodium alginate, Cacl2
  • 32. Organism Trade Name Target Crops Trichoderma virens SoilGard 12G3 Pythium, Rhizoctonia, and Root rots Ornamental and food crop plants grown in greenhouses, Trichoderma harzianum Rifai strain KRL-AG3 Plant Shield® HC Fusarium, Pythium, and Rhizoctonia Cucurbit vegetables, flowers, bedding plants, Trichoderma harzianum Rifai strain KRL-AG2 T-22™ HC Fusarium, Pythium, and Rhizoctonia Agronomic field and row crops, alfalfa, hay and forage crops, bulb crops, cucurbits, fruiting vegetables, herbs, spices, leafy vegetables, cole crops, legumes, root crops, small grains and tuber crops Trichoderma harzianum Rifai strain KRL-AG2 T-22™ Planter Box Fusarium, Pythium, and Rhizoctonia Agronomic field and row crops, alfalfa, hay and forage crops, bulb crops, cucurbits, fruiting vegetables
  • 33.  There are several reputable companies that manufacture government registered products. Trade Name Bio agent Manufacture Eco fit T. viride Hoechst and Schering AgroEvo Ltd, Mumbai India Super visit T. harzianum Fytovita, Czech Republic Soil guard T. virens Certis Inc,Columbia,MD,USA Root pro T. harzianum Efal Agri, Netanyl,Israel Tusal T. Viride +T. harzianum Tusal Carrera Ester, Lleida Spain Agroderma, Bio-cure, Bioderma, Ecofit, Rakshak, Trichosan Trichoderma viride Biocure (B&F) T. Viride and P. flourescens
  • 34. 1.Seed treatment •Dose: @5 g /kg of seed •Method: Make a paste or slurry adding 5 g in 10-20 ml of water . Pour 1kg of seed on to the paste or slurry and mixed properly to coat the seeds uniformly .Shade dry the coated seeds for 20- 30 minutes before sowing 2.Tuber/Rhizome/Cutting treatment: •Dose: @ 10 g /litre of water •Method:Dip the tuber / rhizome/cuttings in the suspension prepare @ 10 g /litre of water. Shade dry for 15 minutes before planting (Medhi,2009)
  • 35. Seedling Treatment: •Dose: 300 g/ha. •Method: Prepare a suspension @ 5-10 g/litre of water. Dip the roots of seedling for 15minutes and shade dry for 15 minutes before transplanting Nursery bed treatment: •Dose: 250 g for 400 sq.m nursery bed. •Method: 1) prepare a suspension by adding 250 g in 50 litres of water and drench the nursery bed soil. •2) Mixed 250 g in 2 kg cow dung / compost/FYM and spread over 400 sq.m. nursery bed and irrigate the bed.
  • 36. Soil treatment: i) Direct broadcasting: •Dose: 300 g/ha. •Method: Mix 300 g in 6 kg of FYM. Broadcast in one ha of land and irrigate the field. ii) Awaited broadcasting •Dose: 30 g/ha •Method: Mixed 30 g in 6 kg of FYM. Cover the mixture with polythene sheet for 7- 15 days and broadcast in the field iii) Furrow application: •Dose:300g /ha. •Method: It is highly effective in root crops like potato, ginger, turmeric etc and sugarcane.The mixture is applied in furrows at the time of earthing up or after 30 days of planting
  • 37. • Decrease disease intensity. • Reduce the use of chemical fungicides. •Reduce undesirable effects from chemical pesticide. • Play a key role in integrated management of diseases •Safe for the users and the farming community. • Provide natural long term immunity to crops and soil ( Shrivastava , 1996 ).
  • 38. •Deterious effects on non-target micro- organisms •Pathogens may develop resistance to the biocontrol agent •Pathogen replacement may follow control of target disease pathogen •Seasonal/weather phenomena can make biocontrol agent ineffective ( Shrivastava , 1996 ).
  • 39. CONCLUSIONS Biological control an attractive alternative to chemicals Trichoderma species are effective in biological control of fungus-induced plant disease Plant diseases cause major loss of food and money Biological control occurs via several mechanisms Competition Antagonism Mycoparasitism