Detailed information regarding ISR in plants by fungal bio control agents

1. welcome
INDUCED SYSTEMIC RESISTANCE IN PLANTS THROUGH FUNGAL
BIOCONTROLAGENTS
Jayappa
Sr. M.Sc (Agri)
PALB 4255
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2. Flow of seminar
o Introduction
o Mechanisms of Fungal Biocontrol Agents(BCF)
o Different Plant responses to BCF’s
o Case studies
o Conclusion
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3. Introduction
• Contact with pathogenic and non-pathogenic microorganisms
activates a broad range of defense mechanisms in plants
• Two main mechanisms are recognized;
1. Systemic acquired resistance
2. Induced systemic resistance
• Biocontrol fungi (BCF) are agents that control plant diseases
which includes Trichoderma spp, Ampelomyces quisqualis,
Paecilomyces lilacinus and others
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4. Host – Pathogen interaction
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5. (Pieterse et al., 2009)11/9/2016 Dept. of Plant Pathology 5

6. Fungal biocontrol (BCF) agents
Bio agent Target diseases / pathogen
Trichoderma spp. Soil borne disease
Cryptococcus albidus Botrytis spp., Penicillium spp.
Gleocladium virens Rhizoctonia solani, Pythium, Phytophthora
Ampelomyces quisqualis Powdery Mildew
Rhizobium leguminosorum Fusarium oxysporum f. sp. ciceris
Glomus mosseae , Paecilomyces
lilacinus
Nematodes
Verticillium lecanii Penicillium digitatum
(Burges et al., 1998)
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7.  Reduce the negative effects of plant pathogens and promote
positive responses in plant.
 Inoculated plants are sensitized to respond more rapidly to
pathogen attack
 Alleviation of abiotic stresses
 Improve photosynthetic efficiency, especially in plants subjected to
various stresses
 Increase nutrients absorption and nitrogen use efficiency in plants
 Enhance the growth and yield parameters
Role of BCF’s
(Shoresh et al., 2010)
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8. Mechanisms of Fungal Biocontrol Agents
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9. Mycoparasitism
(Irina et al., 2011)
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10. “ Interactions that involve a low-
molecular weight compound or an
antibiotic produced by microorganism
that has a direct effect on another
microorganism”
Antibiosis
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11. Fungal biocontrol agents and their metabolites
(Butt et al., 2001)11/9/2016 Dept. of Plant Pathology 11

12. Competition
• Competition for nutrient and space.
• Biocontrol agent decreases the availability of a particular
substance thus limiting the growth of the plant pathogenic agents
Types :
• Exploitation Competition
• Interference Competition
• Preemptive Competition
• Trichoderma spp produce siderophores that chelate iron and stop
the growth of other fungi
(Chet and Inbar ,1994)
 Competitive exclusion
 Coexistence
 Mutual extinction
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13. Fungal compounds involved in induction of plant responses
 Compounds that are released by Trichoderma spp. into the zone of interaction
induce resistance in plants
 Primarily proteins with enzymatic activity
 xylanase, cellulase, swollenin and endochitinase
 Enhance defense, through induction of plant defense–related proteins and
peptides that are active in inducing terpenoid, phytoalexin biosynthesis and
peroxidase activity.
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14. Elicitation of plant to pathogen attack
 Elicitors of plant defense includes
 oligosaccharides
 low-molecular weight compounds
 secondary metabolites
produced by different Trichoderma spp induce expression of pathogenesis
related (PR) proteins when applied to plants and reduce disease symptoms
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15. (Yedidia et al., 2003)
Bacterial cell colonization of leaves sampled from non elicited and
preelicited cucumber plants
Psuedomonas syringae pv.
Lachrymans progresses
towards the inner leaf tissues
mainly by intercellular (IS)
growth
Considerably fewer P. syringae pv.
Lachrymans cells are observed
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16. Systemic induction of defense-related genes and
proteins
• After treatment with T. harzianum, many defense/stress-related proteins
functions were upregulated.
• Stress response enzymes such as
1. Oxalate oxidase and superoxide dismutase (roots)
2. Methionine synthases, glutathione-S-transferase and glutathione dependent
formaldehyde dehydrogenase (FALDH) (shoots)
• They acts as..
Detoxifying enzymes, peroxidase, scavenging enzyme, Heat
shock proteins (stress proteins).
(Yadav et al., 2015)
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17. Gene expression in Trichoderma harzianum T22 Inoculated Maize
up-regulated of defense- and
stress-related genes
Down regulated
( Shoresh and Harman, 2008)
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18. Abiotic stress induced ROS
production and cell death
Alleviation of damage by reactive oxygen species
Equilibrium
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19. Major ROS scavenging antioxidant enzymes
Enzymatic antioxidants Reactions catalyzed
Superoxide dismutase (SOD) O₂⁻ + O₂ ⁻+ 2H+ → 2H₂O₂ + O₂
Catalase (CAT) H₂O₂ ̶ →H₂O + ½ O₂
Ascorbate peroxidase (APX) H₂O₂ + AA →2H₂O + DHA
Guaicol peroxidase (GPX) H₂O₂ + GSH → H₂O + GSSG
Monodehydroascorbate reductase (MDHAR) MDHA + NAD(P)H → AA + NAD(P)+
Dehydroascorbate reductase (DHAR) DHA + 2GSH → AA + GSSG
Glutathione reductase (GR)
GSSG + NAD(P)H → 2GSH +
NAD(P)+
(Gill and Tuteja, 2010)
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20. Perception of the signal and activation of MAPK signaling cascade
Interaction of the signal molecules from the BCF with particular plant receptor
molecules in the interaction zone, activates a MAPK signaling cascade
Plant interaction with Trichoderma results in induction of NBS/leucine rich
repeat resistance protein.
NBS/ LRR are determinants of plant immune system and it trigger a cascade of
signal transduction results in resistance response
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21. (Shoresh et al., 2010)
Cont..
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22. Trichoderma MAMPs currently identified in different species
(Hermosa et al ., 2012)11/9/2016 Dept. of Plant Pathology 22

23. Plant signaling pathways induced by BCF leading to disease resistance
Trichoderma upregulate the Pal1, which encodes for phenylalanine ammonia-lyase
Pal1 is activate JA/ethylene signaling
It catalyzes the first step of phenyl propanoid pathway, leading to production of
phenolic compounds, including phytoalexins
Defense of the plants against infection
(PAL)
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24. Plant growth enhancement by BCF inoculation
Trichoderma species inoculation induces root and shoot growth
The application of Trichoderma lead to an
Increase in dry matter content
Starch and soluble sugars
Germination percentage
 Importantly, the effect of BCF on plant growth has a long duration and
even lasts for the entire life of annual plants
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25. CASE STUDIES
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26. Plant Protection Institute, Hungarian Academy of Sciences, H-1525 Budapest, Hungary
PNAS September 20, 2005 vol. 102 no. 38
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Objective :
To test the effect of P. indica on barley to salt stress tolerance and disease resistance.

27. Impact of Piriformospora indica on salt-stress tolerance and
root infections by Fusarium culmorum.
Plants was determined in final 2 weeks ( totally
5weeks ) in the presence of 0,100 and 300 mM
NaCl, in three independent experiments.
Plant phenotypes demonstrating the protective
potential of P. indica toward F. culmorum.
(Waller et al., 2005)11/9/2016 Dept. of Plant Pathology 27

28. Ascorbate and DHA reductase (DHAR) activity in P. indica-infested roots.
measured in roots of 1-, 2-, and 3-week-old P. indica-infested (shaded
columns) and control (free of P. indica, open columns) barley plants.
(Waller et al., 2005)
ba
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29. Objective:
To prove that suppression of Rhizoctonia solani-incited cotton seedling disease by
T. virens is the result of induction of resistance mechanisms in the cotton host.
Southern Crops Research Laboratory, United States Department of Agriculture-Agriculture
Research Service, 2765 F&B Road, College Station, Phytopathology, 24 November 2000.
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30. Effect of Trichoderma virens on terpenoid concentrations in
cotton roots inoculated with Rhizoctonia solani
HG = Hemigossypol,
dHG = Desoxyhemigossypol,
G = Gossypol
(Howel et al., 2000)
HPLC analyses of seedling radicle extracts for terpenoid content
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120.0022.6239.79

31. Assay of the biocontrol efficacy of strains of T. virens, T. koningii
and T. harzianum
1. T. virens strains : G-6, G-11, G6-5, G-4, and GTH-34
2. T. harzianum strain TH-23 ,GTK-53 and GTK-56
3. T. koningii strain TK-7, the T. virens mutant strain G6-4, and WB+PM
control
Significant reduction
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32. Objective :
To evaluate the potential of fungal bio control agent, Trichoderma harzianum to
control the root-knot nematode Meloidogyne javanica.
Department of Nematology, Agricultural Research Organization (ARO), The Volcani Center,
P.O.B. 6, Bet- Dagan 50250, Israel; The American Phytopathological Society, Vol. 91, No. 7,
2001.
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33. Effect of Trichoderma harzianum on Meloidogyne javanica
Green house condition
Natural condition
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34. Evaluation of the activity of Trichoderma proteinase Prb1-
transformed lines
Non treated control (C)
Wild-type (WT)
Prb1-transformed strains
P-1, P-2, P-5, and P-6
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35. Objectives :
• To test Growth enhancement ( increased root production) in treated plants
• The mechanisms of control of the seed and root pathogen Pythium ultimum
and the foliar pathogen Colletotrichum graminicola in maize
Departments of Horticultural Sciences and Plant Pathology, Cornell University, Geneva, NY
14456, The American Phytopathological Society, Vol. 94, No. 2.
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36. Interactions Between Trichoderma harzianum Strain T22 and Maize
Inbred Line Mo17
Seedlings of maize line Mo17 (10 days old) grown with or without a seed
treatment with T22
( Harman et al., 2004)11/9/2016 Dept. of Plant Pathology 36

37. Plant height and stalk diameter are greater in the presence of T22.
Eight week old plants of maize line Mo17
Cont’d…,
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38. Effect of T22 on Pythium ultimum
(Harman et al., 2004)
5 day old Maize inbred line Mo17 in 2 separate experiment
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39. Effect of T22 on C. graminicola
More diseased
and chlorotic
leaf area control
Seed treated with T22
(Harman et al., 2004)
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40. Advantages
• More sustainable method of crop production
• Reduce environmental pollution
• Increased biodiversity
• More target specific than chemicals
• Consequently, plants treated with beneficial fungi will be larger and
healthier and have better yields than plants without them.
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41. Conclusion
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