Trichoderma march 14th

76
W E L C O M E

description

 

Transcript of Trichoderma march 14th

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WELCOME

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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 .

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Use of Trichoderma in plant disease management

KITTUR RANI CHANNAMMA COLLEGE OF HORTICULTURE, ARABHAVI - 591 218

University of Horticultural Sciences, Bagalkot

Presented by:PRADNYARANI P. N

USH11PGM135DEPT. OF

HORTICULTURE PLANT PATHOLOGY

Seminar II

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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.

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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

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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 

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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

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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

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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

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

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Conidiophores

Conidia

Hyphae

Morphological structure of Trichoderma

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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).

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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

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

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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

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Hypocrea teliomorph of Trichoderma spp.

1mm Mature perithecia

20 μmViewed through stereo microscope Samuels et al., 2006, USA

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Where do they come from?

They can be easily isolated from soil, root, decaying wood and other forms of plant organic matter

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Singh et al., 2007.

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enzyme production

Antibiotic production

Mycoparasitism

SAR

Competition

Growth promotion

Effective antagonist

Rapid substrate colonization

Potential bio control activities exhibited by Trichoderma

Kamala and Indira, 2012, Manipur

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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

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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

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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).

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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

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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.

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Growth inhibition of R. solani by the T. virens produced antibiotic gliotoxin . A: Gliotoxin amended B: non amended

Cont…

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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

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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…

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Evaluation under in vitro techniques

Dual culture or paired culture

Upadhyay and Rai (1987)

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Filter paper disc method

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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)

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Mass production of biocontrol agentsLiquid 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

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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

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• 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

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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

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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.

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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

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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.

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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

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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.

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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

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Case studies

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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

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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

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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

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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 14 th 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

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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

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Storage days

Agro-wastes/by-products(106 cfu g-1)

Sand maize meal

Rice husk

Saw dust Groundnut 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

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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

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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

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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

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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

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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

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Treatement Plant height(cm) Plant dryWeight(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%

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

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

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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

30 c⁰ 29.15 17.2 43.67 35.41 36.89 31.37

25 c⁰ 36.04 38.89 37.56 36.44 35.3 30.96

20 c⁰ 31.14 26.89 39.41 33.63 31.67 29.96

*Values are means of four replicates.

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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

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Fig8: Inhibitory effect of the culture filtrate of Trichoderma spp. Incubated at different temperature (5, 15, 25, 35, 40 C). Each value is an average of three ⁰replicates.T1s= Trichodema harzianum isolate T1s, TvPD = T. viride isolate TvPD, TDPs = T. harzianum isolate TDPs Perveen and Bokhari, 2012, Saudi Arabia

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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

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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 ± SEMValues in the column followed by same letter are not significantly different (P<0.05).

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Fig10: Efficiency of P solubility and biocontrol activity of T .harzianum isolates against Xanthomonas sp.

Padmavathi and Madhumathi, 2011, Banglore

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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

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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

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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

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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

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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

Sclerotioalcount

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

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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

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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

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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

Page 70: Trichoderma march 14th

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.

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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

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Healthy egg ofHeterodera glycines

Egg parasitizedBy fungus

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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

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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

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CONCLUSION

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.

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THANKU