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Transcript of Trichoderma march 14th
WELCOME
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
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
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.
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
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
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
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
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
Fig. 1 (A) Trichoderma on solid media (B) microscopic view (C) Trichoderma in liquid medium
Conidiophores
Conidia
Hyphae
Morphological structure of Trichoderma
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).
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
Fig3 : View of T. harzianum through a Stereo microscope (1mm to 10μm) Samuels et al., 2006, USA
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
Hypocrea teliomorph of Trichoderma spp.
1mm Mature perithecia
20 μmViewed through stereo microscope Samuels et al., 2006, USA
Where do they come from?
They can be easily isolated from soil, root, decaying wood and other forms of plant organic matter
Singh et al., 2007.
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
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
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…
Evaluation under in vitro techniques
Dual culture or paired culture
Upadhyay and Rai (1987)
Filter paper disc method
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)
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
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
• 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
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
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.
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
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.
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
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.
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
Case studies
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
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
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
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
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
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
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
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
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
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
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
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%
Fig5: Pathogen growth inhibition by Trichoderma after 6 day of inoculation in dual culture Hajieghrari et al., 2008, Iran
Fig6: Pathogen growth inhibition Trichoderma volatile compounds Hajieghrari et al., 2008, Iran
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.
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
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
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
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).
Fig10: Efficiency of P solubility and biocontrol activity of T .harzianum isolates against Xanthomonas sp.
Padmavathi and Madhumathi, 2011, Banglore
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
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
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
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
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
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
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
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
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.
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
Healthy egg ofHeterodera glycines
Egg parasitizedBy fungus
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
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
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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