1. W
E
L
C
O
M
E

2. In agriculture, world wide, pathogens are threat to crop production
(Sarah and Paul, 2005)
 The extensive use of fungicides in various parts of the world for years has
increased the pollution level in soil and water, and adverse effect on food
quality and human health
 Apart from this, the chemicals tend to become less efficient due to the
development of resistance among the pathogen a over time
Hence, it is necessary to look for alternative disease management
practices, which include the use of eco-friendly biological control agents .

3. KITTUR RANI CHANNAMMA COLLEGE OF HORTICULTURE,
ARABHAVI - 591 218
Presented by:
PRADNYARANI P. N
USH11PGM135
DEPT. OF
HORTICULTURE PLANT
PATHOLOGY
3

4. What are BIO-CONTROL AGENTS ?
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) defn:- “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.

5. Trichoderma is….
 Very effective biological agent
 Free living
 Ubiquitous
 Highly proliferating
 Non- pollutive
 Easily accessible
 Non phytotoxic
 Systemic ephemeral
 Readily biodegradable
 Cost effective
 Synergistic effect
 Longer shelf life
 Greater compatibility

6. History of Trichoderma
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

7. Trichoderma
 Free living fungus common in soil
and root ecosystem
 Highly interactive in root, soil and
foliar environment
 Suppresses the pathogen by
different mechanism of biocontrol
Trichoderma harzianum

8. Taxonomical position 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

9. General Characters of Trichoderma spp.
 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

10. Fig. 1 (A) Trichoderma on solid media (B) microscopic view (C) Trichoderma
in liquid medium

11. Conidiophores
Conidia
Hyphae
Morphological structure of Trichoderma

12. Fig 2: Photograph shows colonies of Trichoderma strains on PDA plate (dorsal
view) and conidiophore with conidia. 1a & 1b. T. virens (IMI-392430), 2a & 2b.
T. pseudokoningii (IMI-392431), 3a & 3b. T. harzianum (IMI-392432), 4a &4b.
T. harzianum (IMI- 392433) and 5a & 5b.T. harzianum (IMI-392434).

13. Conidiophores characteristics of
Trichoderma spp.
 Highly branched, difficult to define
or measure
 Loosely or compactly tufted
 Main branches of the
conidiophores produce lateral side
branches
 The branches may rebranch, with
the secondary branches and longest
secondary branches being closest to
the main axis

14. Fig3 : View of T. harzianum through a Stereo microscope (1mm to 10μm)
Samuels et al., 2006, USA

15. Conidia
 Typically appear dry but in
some species they may be held
in drops of clear green or
yellow liquid (e.g. T. virens, T.
flavofuscum)
Round to oval in shape

16. Hypocrea teliomorph of Trichoderma spp.
1mm
Mature perithecia
20 μm
Viewed through stereo microscope Samuels et al., 2006, USA

17. Where do they come from?
They can be easily isolated from soil, root, decaying
wood and other forms of plant organic matter

18. Singh et al., 2007.

19. enzyme
production
Antibiotic
production
Mycoparas
itism
SAR
Competition
Growth
promotion
Effective
antagonist
Rapid
substrate
colonization
Potential bio control activities exhibited by Trichoderma
Kamala and Indira, 2012, Manipur

20. Competition
 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…

28. Evaluation under in vitro techniques
 Dual culture or paired culture
Upadhyay and Rai (1987)

29. Filter paper disc method

30. Procedure for isolation of Trichoderma from soil
Isolation from soil on selective medium
incubate 7 days at 250 C
Sub culturing on PDA plates
Purification
Inoculation of purified culture on PDA
slants
Preservation in deep freezer (-200 C)

31. Mass production of biocontrol agents
Liquid fermentation method
Mix 30 gm molasses and 6gm Brewer’s yeast in 1
litre of water. Distribute 60 ml in each conical flask.
autoclave
Inoculate 8mm mycelial discs of Trichoderma in
medium
Incubate for 10 days at room temperature
Use for multiplication in the fermentor
Prepare 50 lit of molasses + yeast medium and
sterilize for 30 min in the fermentor
Transfer aseptically 1 lit of Trichoderma

32. Incubate for 10 days
using haemocytometer (108 /ml spore)
500 ml of fungal biomass + 1 kg of
talc powder
Air dry & and carboxy methyl
cellulose (CMC) + sticker 5 gm / kg
Store in polythene bag

33. • 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

34. Advantages
 Enhances yield along with quality of produce
 Boost germination rate
 Increase in shoot & Root length
 Solubilising various insoluble forms of Phosphates
 Augment Nitrogen fixing
 Promote healthy growth in early stages of crop
 Increase Dry matter Production substantially

35.  Harmless to humans and livestock
 Act against a wide range of pathogenic fungi
 Perpetuate themselves by producing ample spores
 Grow rapidly and quickly colonize the soil
They can promote nutrient uptake and enhance plant growth
Provide natural long term immunity to crops and soil.

36. Disadvantages
• Harmful parasite of mushrooms
• Looses its effectivity if not placed in its native condition.
• It cannot be used as foliar spray
• It do not grow in alkaline pH (above 8).
• Zone specific & slow growth

37. Methods of application
1. Seed treatment: Mix 6 - 10 g of Trichoderma powder per Kg of seed before sowing.
2. Nursery treatment: Apply 10 - 25 g of Trichoderma powder per 100 m2 of nursery
bed. Application of neem cake and FYM before treatment increases the efficacy.
3. Cutting and seedling root dip: Mix 10g of Trichoderma powder along with 100g of
well rotten FYM per litre of water and dip the cuttings and seedlings for 10 minutes
before planting.

38. 4. Soil treatment: Apply 5 Kg of Trichoderma powder per ha after turning of sun
hemp or dhaincha into the soil for green manuring Or Mix 1kg of Trichoderma
formulation in 100kg of farmyard manure and cover it for 7 days with
polythene. Sprinkle the heap with water intermittently. Turn the mixture in
every 3-4 days interval and then broadcast in the field.
5. Plant Treatment: Drench the soil near stem region with 10g Trichoderma
powder mixed in a litre of water
6. Wound application
7. Furrow application
Kulkarni and Sagar, 2007

39. Precautions
Don't use chemical fungicide after application of Trichoderma
for 4-5 days.
Don't use Trichoderma in dry soil. Moisture is a essential
factor for its growth and survivability.
Don't put the treated seeds in direct sun rays.
Don't keep the treated FYM for longer duration.

40. Compatibility
Compatible with Organic manure, biofertilizers like
Rhizobium, Azospirillum, Mycorrhizae, Azotobacter, Bacillus
Subtilis and Phosphobacteria, Gliocladium virens, Pseudomonas
fluorescens
Trichoderma can be applied to seeds treated with Metalaxyl or
Captan, Carboxin, Carbendazium but not Mercurials.
Kulkarni and Sagar, 2007

41. Case studies

42. Fig1: Incidence of F. oxysporum f.sp. cepae in onion sets raised from seeds treated with
procholaz and Trichoderma harzianum in artificially pathogen-inoculated pot soil.
C+: sets raised from non-treated seeds in pot soil inoculated with FOC16. C-: sets
raised from non treated seeds in non inoculated pot soil. Bars topped by the same
letter do not differ significantly according to the Tukey-Kramer test at P<0.05
Coskuntuna and Ozer., 2008, Turkey

43. Fig2: Incidence of F.oxysporum f.sp. cepae sets raised from seeds treated with
prochloraz and Trichoderma harzianum in naturally pathogen infested field
soil. Control sets raised from seeds in field soil infested with FOC. Bars topped
by the same letter do not differ significantly according to the Tukey-Kramer test
at P<0.05.
Coskuntuna and Ozer., 2008, Turkey

44. Fig3: Antagonistic activity of Trichoderma against different pathogens. A. R.
solani. B. F. oxysporum, C. P. ultimum and D. P. aphanidermatum
Kamala and Indira., 2012, Manipur

45. Fig4: Variation of the ratio of late blight infected leaves with respect to the leaf position
on the main stem. The data were taken at the 14th day after the foliar inoculation
of P. infestans. The values calculated as ratio of infected to total leaves on the
main stem. Zegeye et al., 2011, Ethiopia

46. Storage
days
Talc Vermicompost Mean
Room
temperature
Refrigerator
temperature
Mean Room
temperature
Refrigerator
temperature
Mean
30 143.33 132.00 137.67 186.00 147.00 166.50 152.08
60 113.00 89.00 101.00 187.00 89.00 138.00 119.50
90 98.00 77.00 87.50 152.00 76.33 114.17 100.83
120 68.00 64.00 66.00 108.00 41.33 74.67 70.33
150 46.00 39.00 42.50 51.00 27.33 39.17 40.83
180 28.00 25.00 26.00 24.00 16.00 20.00 23.25
Mean 82.72 71.00 76.86 118.00 66.17 92.08 84.47
Table1: Effect of different carrier materials on shelf life ( 106 cfu g -1 ) of T. harzianum (Th-2)
at Different temperatures Bheemaraya et al., 2011, Raichur
Comparing of means C.D @ 1%
Carrier(A) 2.005
Temperature(B) 2.005
Storage days(C) 3.472
AXB 2.835
AXC 4.911
BXC 4.911
AXBXC 6.945
Room Temperature=28 1oc
Refrigerated temperature=4 1oc

47. Storage
days
Agro-wastes/by-products( 106 cfu g-1)
Sand maize
meal
Rice
husk
Saw dust Groundn
ut cake
Castor
cake
Mean
30 39.00 65.67 3.70 32.33 38.00 35.74
60 38.67 64.00 3.67 31.00 35.67 34.60
90 38.00 62.33 3.53 30.33 34.67 33.77
120 5.63 53.00 2.97 25.67 24.00 22.25
150 0.64 1.27 2.23 9.67 14.67 5.69
180 0.53 0.70 1.63 7.67 7.83 3.67
Mean 20.41 41.16 2.96 22.78 25.81 22.62
Table2: Effect of different agro-wastes/by-products on shelf life of Trichoderma piluliferum (Tp)
Bheemaraya et al., 2011, Raichur
C.D.at 1%
Agro wastes(A) 0.439
Storage days(B) 0.481

48. Bio-agents Percent inhibition of mycelial growth*
T. viride
(TV-3)
T. harzianum
(TH-2)
T. piluliferum
(TP)
B. subtilis (E) 100.00
(89.99)**
0.00
(0.00)
0.00
(0.00)
P. flourescens (I)
(Pf-4)
100.00
(89.99)
0.00
(0.00)
0.00
(0.00)
A. quisqualis (E) 10.67
(18.63)
0.00
(0.00)
10.00
(18.42)
control 0.00
(0.000
0.00
(0.00)
0.00
(0.00)
mean 52.66
(49.65)
0.00
(0.00)
2.50
(4.6)
C.D.at 1% 1.10 NS 0.69
Table3: Effect of different bio-agents on compatibility of Trichoderma spp.
Bheemaraya et al., 2011, Raichur

49. Treatment Percent inhibition of mycelial growth* Mean
Concentration(%)
2.5 5.0
T1- NSKE 0.00
(0.00)**
0.37
(2.02)
0.19
(1.01)
T2 -Nimbicidine 81.48
(64.51)
83.70
(66.19)
82.59
(65.35)
T3- Prosophis leaf extract 48.15
(43.93)
57.04
(49.04)
52.59
(46.49)
T4- Pongamia leaf extract 0.37
(2.02)
10.37
(18.78)
5.37
(10.40)
T5- Eucalyptus leaf
extract
0.00
(0.00)
0.00
(0.00)
0.00
(0.00)
T6- control 0.00
(0.00)
0.00
(0.00)
0.00
(0.00)
Mean 21.67
(18.41)
25.25
(22.67)
23.46
(20.54)
Table4: Effect of different plant extracts on compatibility of Trichoderma harzianum (Th-2)
Bheemaraya et al., 2011, Raichur
S.EM C.D.at 1%
Plant extracts (P) 0.60 2.35
Concentration (C) 0.34 1.36

50. Treatment Percent inhibition of mycelial growth* Mean
Concentration(%)
0.1 0.2
T1- Mancozeb 0.37
(2.02)**
5.19
(13.14)
2.78
(7.58)
T2 - Carbendazim 100.00
(89.99)
100.00
(89.99)
100.00
(89.99)
T3- Captan 100.00
(89.99)
100.00
(89.99)
100.00
(89.99)
T4- Propiconozole 100.00
(89.99)
100.00
(89.99)
100.00
(89.99)
T5- Metalaxyl-
m+mancozeb
1.85
(4.54)
11.48
(19.59)
6.67
(12.07)
T6- Control 0.00
(0.00)
0.00
(0.00)
0.00
(0.00)
Mean 50.37
(46.09)
52.78
(50.45)
51.57
(48.27)
Table5: Effect of different fungicides on compatibility of Trichoderma harzianum (Th-2)
Bheemaraya et al., 2011, raichur
S.EM C.D.at 1%
Fungicide (F) 1.12 4.42
Concentration (C) 0.65 2.55

51. Treatment Percent inhibition of mycelial growth* Mean
Concentration(%)
0.1 0.2
T1-Chloropyriphos 96.30
(83.50)
100.00
(89.99)
98.15
(86.74)
T2 –Carbofuron 100.00
(89.99)
100.00
(89.99)
100.00
(89.99)
T3- Indoxocarb 87.78
(69.69)
100.00
(89.99)
93.89
(79.84)
T4- Imidachloprid 12.96
(21.09)
59.26
(50.33)
36.11
(35.71)
T5- Control 0.00
(0.00)
0.00
(0.00)
0.00
(0.00)
Mean 59.41
(52.85)
71.85
(64.06)
65.63
(58.46)
Table6: Effect of different insecticides on compatibility of Trichoderma harzianum (Th-2)
Bheemaraya et al., 2011, Raichur
S.EM C.D.at 1%
Insecticides (I) 1.53 6.14
Concentration (C) 0.96 3.88
Mean of four replications, **Figures in parentheses are arcsine transformed values

52. Table7: Effect of soil treatment with formulated Trichoderma species on incidence of
fusarium wilt disease of giza 3 bean cultivar under greenhouse and field conditions.
Nashwa et al., 2008, Egypt
Wilt rating under green house conditions
TREATMENT
Time of application TH 1** TV 1 TS 3 infected control Mean
Two weeks before planting 3.7 4.1 5.2 8.0 5.3
At time of planting 2.5 3.1 4.0 7.0 4.2
Mean 3.1 3.6 4.6 7.5
L.S.D at 0.05 time of application (A ):0.5 bioagents (B) :0.43. Interaction (AXB) :0.61
Wilt rating * under field conditions
Time of application TH 1 TV1 TS 3 Infected control Mean
Two weeks before planting 3.3 4.2 5.0 6.0 4.7
At time of planting 4.0 4.2 5.0 7.0 5.1
Mean 3.6 4.1 5.0 6.5
L.S.D at 0.05 time of application a :0.2 bioagents b :0.25. Interaction (AXB) :0.36
•According to CIAT scale (van schoonhoven and pastor-corrales,1987)
TH 1: T. harzianum, TV 1: T. viride, TS 3: T. virens

53. Treatement Plant height(cm) Plant dry
Weight(g)
Disease incidence
C 14.52a 2.45a 0.00a
Fs 6.52b 1.06b 1.80b
Fs+ Tv 12.68c 2.42a 1.06c
Fs+ Th 14.18d 2.58a 0.80d
Table8: Effect of T. harzianum and T. viride on height.dry weight and disease
incidence of tomato plants inoculated with F. Solani under pot conditions.
Perveen and Bokhari., 2012, Saudi Arabia
Each value is average of six replicates. Data Followed by different letters in the column are
significantly different (p< 0.05 ) according to Duncan’ s multiple range test .
C, uninoculated control
Fs ,F solani :
Tv: T . viride ,
Th: T. harzianum
Disease incidence graded on 0 to 3 scale where ,0= 25 %severity , 1 =26 to 50%,
2 = 51 to 75% and 3= 76 to 100%

54. Fig5: Pathogen growth inhibition by Trichoderma after 6 day of inoculation in dual
culture Hajieghrari et al., 2008, Iran

55. Fig6: Pathogen growth inhibition Trichoderma volatile compounds
Hajieghrari et al., 2008, Iran

56. Table9: Effect of pH and temperature on the mycelial growth(mm) of Trichoderma isolates
Hajieghrari et al., 2008, Iran
Treatment T. hamatum
T612
T. harzianum
T447
T. virens
T523
T. harzianum
T969
Trichoderma
sp. T
T. hamatum
T614
pH 8 33.26* 23.89 41.15 31.07 30.19 33.78
pH 7 29.3 24.3 33.96 39.45 30.45 30.85
pH 5 33.78 34.8 45.52 34.7 32.22 27.68
30⁰c 29.15 17.2 43.67 35.41 36.89 31.37
25⁰c 36.04 38.89 37.56 36.44 35.3 30.96
20⁰c 31.14 26.89 39.41 33.63 31.67 29.96
*Values are means of four replicates.

57. Fig7 :Antagonistic activity of Trichoderma species against F.oxysporum evaluated by
dual culture interaction
(A)F.oxysporum alone ,
(B) F oxysporum +T.harzianum (T 1s),
(C) F.oxysporum + T.viride (TvPDs)
(D) F.oxysporum + T.harzianum(TDPs) Perveen and Bokhari, 2012, Saudi Arabia

58. Fig8: Inhibitory effect of the culture filtrate of Trichoderma spp. Incubated at different
temperature (5, 15, 25, 35, 40⁰C). Each value is an average of three replicates.T1s=
Trichodema harzianum isolate T1s, TvPD = T. viride isolate TvPD, TDPs = T.
harzianum isolate TDPs Perveen and Bokhari, 2012, Saudi Arabia

59. Fig9: Average liner growth rate (ALG) of Trichoderma species on various culture media. Each value is
an average of three replicates. T1s= Trichoderma harzianum isolate T1s, TvPDs= T. viride isolate
TvPDs, TDPs=T harzianum isolate TDPs, PDA=Potato dextrose agar, SDA =Sabouraud dextrose
agar, WA = Water agar (2% agar), CDA = Czapek dox agar, PDAL= natural media agar (PDA + 1%
date palm leaves)
Perveen and Bokhari, 2012, Saudi Arabia

60. Table10 : Effect of strains of Trichoderma species on the per cent inhibition of radial
colony growth of P. aphanidermatum Mishra, 2010, India
Trichoderma species Percent inhibition
T.harzianum -4532 60.3 0.3 e
T harzianum-4572 69.8 0.3 g
T.viride -801 54.1 0.5 c
T.viride-1763 52.2 0.5 b
T.viride-1433 72.0 0.3 h
T.viride-793 62.1 0.3 f
T.Viride-2109 50.4 0.4 a
T.koningii-2385 56.4 0.2 d
T.virens-2023 53.5 0.6 c
T.virens-2194 59.6 0.6 e
Values are average of three replicates SEM
Values in the column followed by same letter are not significantly
different (P<0.05).

61. Fig10: Efficiency of P solubility and biocontrol activity of T .harzianum isolates
against Xanthomonas sp.
Padmavathi and Madhumathi, 2011, Banglore

62. Fig11: Effect of initial pH on chitinase and ß-1,3-glucanase production (using
0.5% chitin or laminarin as carbon source, respectively ) by T.harzianum
Katatny et al., 2000, Egypt

63. Fig12: Effect of different carbon sources on chitinase and ß-1,3-glucanase production by T.
harzianum and on inhibition of S.rolfsii (100 % ß-1,3-glucanase activity correspond to
14.7 nkat/mL and 100% chitinase activity correspond to 59.8 pKat/mL)
Katatny et al., 2000, Egypt

64. Fig13: Release of total reducing sugars(R,S),glucose and N-acetyl glucosamine
from S.rolfsii(dried and fresh mycelium),T.harzianum and chitin by the
T.harzianum enzymes Katatny et al., 2000, Egypt

65. Table11: Evaluation of Trichoderma isolates against soil borne fungal pathogens using dual
culture Amin et al., 2010, Jammu and Kashmir
Treatment Radial growth (mm)of test pathogens
R.solani S.rolfsii S.sclerotiorum
Trichoderma virens(Ts-1) 46.55
(48.11)
36.26
(59.71)
56.19
(37.56)
Trichoderma harzianum(Th-1) 35.43
(60.51)
34.67
(61.47)
39.08
(56.57)
Trichoderma harzianum(Th-2) 43.32
(51.71)
35.33
(60.75)
55.69
(38.12)
Trichoderma viride(Tv-1) 30.67
(65.71)
28.88
(67.91)
30.41
(66.21)
Trichoderma viride(Tv-2) 25.65
(71.41)
32.00
(64.44)
34.28
(61.91)
Trichoderma viride(Tv-3) 41.59
(53.64)
34.93
(61.18)
55.65
(38.16)
Control 89.72 90.00 90.00
C.D.(P=0.05) 2.52 1.23 3.59
Figures in parenthesis are per cent inhibition values

66. Table12: Evaluation of Trichoderma isolates against production of sclerotia in soil fungal
pathogens using dual culture Amin et al., 2010, Jammu and Kashmir
Treatment Rhizoctonia solani Sclerotrum rolfsii Sclerotinia sclerotiorum
Sclerotioal
count
Inhibition
over
control(%)
Sclerotial
count
Inhibition
over
control(%)
Sclerotial
count
Inhibition over
control(%)
(Ts-1) 35.59 66.63 38.66 67.60 19.09 39.70
(Th-1) 23.66 77.81 28.73 75.92 12.07 61.87
(Th-2) 31.73 70.25 34.29 71.26 18.12 42.76
(Tv-1) 17.33 83.75 23.64 80.18 9.45 70.15
(Tv-2) 19.47 81.75 26.07 78.15 11.12 64.87
(Tv-3) 27.25 74.45 33.78 71.69 15.57 50.82
Control 106.66 - 119.3 - 31.66 -
C.D.(P=0.05) 1.89 2.07 0.98

67. Fig14: The level of hydrolytic enzymes activities from three different samples E2-
extracts from Botrytis mycelia;E3-extracts from Trichoderma mycelia :E4:Extract
from a mixture of pathogen and antagonistic strains
Cornea et al., 2009, Romania

68. Table13: Evaluation of volatile metabolites produced by Trichoderma isolates against
production of sclerotia in different pathogens
Amin et al., 2010, Jammu and Kashmir
Treatment R. solani S. rolfsii S. sclerotiorum
Sclerotial
count
Inhibition
over control
(%)
Sclerotial
count
Inhibition
over control
(%)
Sclerotial
count
Inhibition
over
control(%)
(Ts-1) 60.09 39.30 91.43 29.30 10.12 57.24
(Th-1) 42.67 56.89 75.66 41.49 6.11 74.18
(Th-2) 54.67 44.77 89.06 31.13 8.73 63.11
(Tv-1) 38.42 61.19 67.00 48.19 5.00 78.87
(Tv-2) 34.00 65.65 71.04 45.07 5.11 78.41
(Tv-3) 49.52 49.97 84.93 34.33 7.22 69.49
Control 99.00 - 129.33 - 23.67 -
C. D
(P=0.05)
3.13 4.76 0.91
Figures in parenthesis are percent inhibition values

69. Fig16:Compatibility test between T. viride and P. fluorescens. The picture was
taken on the 9th day after dual inoculation.
Zegeye et al., 2011, Ethiopia

70. Table14 : Effect of foliar application of T.viride and P.fluorescens on the progress of
late blight disease of potato Zegeye et al., 2011, Ethiopia.
Treatments Mean AUDPC
T. viride 260.0 190.0 c
P. fluorescens 765.1 218.6 b
Mixed culture 999.0 274.5 a
Mancozeb 85.9 77.8 cd
Negative
control(inoculated/untreated)
1045.1 227.2 a
Positive control (non-
inoculated/untreated)
0.00 0.00 d
Means followed by the same letter are not significantly different.
The AUDPC was calculated from five consecutive weekly assessment of
percentage of leaf area with symptoms of late blight. The nine replicates were
arranged in a CRB design and the midpoint rule was used to calculate AUDPC
values.

71. Mechanism of action against
Phytonematodes
• Secretion of Lytic enzyme chitinase help parasitism of
Meloidogyne and Globodera eggs
• T. viride releases Dermadin helps in destruction of
nematode cuticle
• Trichoderma spp. have high rhizosphere competency and
easily colonize the roots, reduce the feeding sites for
nematodes
Jonathan, 2010, New Delhi

72. Healthy egg of
Heterodera glycines
Egg parasitized
By fungus

73.  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

74. Formulations
 Powder formulations
 Encapsulation in organic polymer like sodium alginate
 As spray from emulsifiable concentrates
 Molasses enriched clay granules
 Pellating biomass and bran with sodium alginate

75. The use of Trichoderma has gained importance in managing most of the
plant pathogens.
However, there is still considerable interest in finding more efficient
mycoparasitic fungi especially within Trichoderma harzianum strains,
which differ with respect to their biocontrol effectiveness.
The technique for mass production and use of these bio agents have been
commercialized for the purpose of producers and farmers.

76. U