Role of Induced Systemic Resistance (ISR)In Plant Disease Management

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Role of Induced Systemic Resistance (ISR) In Plant Disease
Management
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Welcome
to PG
Seminar
Series
2019-20

Role of Induced Systemic Resistance (ISR)
In Plant Disease Management
Course No.: Pl. Path. 591
Speaker
Vaniya Ravikumar G.
Reg No: 2010118141
3rd Sem. M. Sc. (Agri.) Plant Pathology
Major Advisor
Dr. Pushpendra Singh
Associate Professor
Dept. of Plant Pathology
College of Agriculture
NAU, Waghai
Co - guide
Dr. J. J. Pastagia
Associate Professor
Dept. of Agril. Entomology
College of Agriculture
NAU, Waghai
2

Content
Introduction
History of ISR
Characteristics of ISR
Mechanisms of SAR
Types of ISR
Different PGPRs elicited ISR
Case studies
Conclusion
3
JA and Ethylene Biosynthesis pathway

Crop losses caused by pathogens, animals and weeds are all
together responsible for losses ranging between 20 to 40 per cent of
global agricultural productivity (Oerke, 2006).
During 2016-17, total area under cereals cultivation in India was
99.34 million hectares with production of 242.70 million tonnes
(Anonymous, 2017).
As per National Horticulture Database published by National
Horticulture Board, during 2016-17, India produced 92.846 million
metric tonnes of fruits and 175.008 million metric tonnes of vegetables
(Anonymous, 2017).
The area under cultivation of fruits stood at 6.480 million hectares
while vegetables were cultivated at 10.290 million hectares.
Introduction
www.agricoop.nic.in
http://nhb.gov.in/statistics/Publication/Horticulture 4

Fig.1. Abiotic and biotic factors causing crop losses
Oerke (2005), Crop losses to pests, Journal of Agricultural science, 31-43. 5

Plant disease control is mainly based on the uses of fungicides,
bactericides, and insecticides, chemical compounds toxic to plant
invaders, causative agents, or vectors of plant diseases. However, the
detrimental effect of these chemicals on the environment and human
health is unavoidable.
Therefore, it is essential to introduce environmentally friendly
alternative measures for management of plant diseases. Induced plant
resistance is one of the promising non-chemical strategies for the
effective management of diseases.
Induced Systemic Resistance (ISR) is a resistance mechanism
in plants that is activated by infection. Its mode of action does not
depend on direct killing or inhibition of the invading pathogen, but
rather on increasing physical or chemical barrier of the host plant.
6

Induced Systemic Resistance (ISR) signal
transduction pathways activated through jasmonate and ethylene.
Induced systemic resistance has been induced by Abiotic and
biotic factors. In abiotic factors such as temperature, radiations and
other chemicals may induce a number of defense compounds in
plants. And biotic factors including different bio agents viz.,
Trichoderma sp., Pseudomonas sp., Bacillus sp., PGPR (Plant
Growth Promoting Rhizobacteria) are responsible for plant
induced resistance.
Plants protect themselves against biotic factors by physical
strengthening of the cell wall, and producing various PR proteins
including defense-related enzymes such as Peroxidase, β-1,3-
glucanase, Chitinase, Phenylalanine ammonia lyase and
Polyphenol oxidase and Phenol content in response to pathogen
infection.
7

The earliest reports of what appears to be induced resistance
to disease come from the late nineteenth century and the first part of
the twentieth century.
For example, Ray and Beauverie independently reported that
attenuated strains of Botytis cinerea induced resistance to that same
pathogen (Chester, 1933). In addition, Ray also found that treatment
of several plant species with attenuated Botrytis or extracts of the
pathogen resulted in enhanced resistance against soft rot bacterium
referred to as ‘Bacillus putrefaciens’.
In 1940, Muller and Borger reported that prior inoculation
of the cut surface of a potato tuber with an avirulent race of
Phytophthora infestans resulted in the local induction of resistance to
virulent races of the same pathogen.
8
History

In 1960, Cruickshank and Mandryk found that injecting stems of
tobacco plants with sporangia of Personospora tabacina induced resistance
in the foliage to further infection by the same pathogen.
In 1975, Kuc and colleagues found that droplet inoculation of one
leaf of anthracnose susceptible cucumber with the cucumber anthracnose
fungus Colletotrichum orbiculare induced systemic resistance to the same
pathogen.
Wei et al. (1991) and Van Peer et al. (1991) reported that
resistance could be induced in cucumber and carnation, respectively, by
PGPR (Plant Growth Promoting Rhizobacteria).
Uknes et al. (1992) were the first to demonstrate biologically
induced resistance in Arabidopsis by inducing resistance to Turnip crinkle
virus (TCV) and Pseudomonas syringae pv. tomato (Pst) by prior
inoculation of the plants with necrosis inducing TCV.
https://onlinelibrary.wiley.com/doi/pdf/10.1002/9781118371848.ch1
9

Induced Systemic Resistance (ISR)
Induced systemic resistance (ISR) emerged as an
important defense mechanism by which selected plant growth
promoting bacteria (PGPR) and fungi in the rhizosphere primed
the entire plants for enhanced defense against a number of plant
pathogens.
ISR is elicited by a local infection, plants respond
with a jasmonic acid and ethylene dependent signalling cascades
that lead to the systemic expression of a broad spectrum and long-
term disease resistance which is effective against fungi, bacteria
and viruses.
ISR is the activation of latent innate immune
responses. It develops systemically in response to colonization of
plant root by PGPR and mycorrhizal fungi.
10

ISR does not involve the accumulation of pathogenesis-
related proteins or but instead, relies on jasmonic acid and ethylene
dependent signalling pathways.
ISR is a phenomenon where plant/s treated with certain
chemicals or inoculated with pathogen avirulent strain produce a
signal compound that is transported systemically throughout the plant
and activates its defense mechanism without its own physical
presence at the site.
Plant resistance to pathogens and pests can be active and/or
passive (Hammerschmidt and Nicholson, 1999).
Passive resistance depends on defenses that are constitutively
expressed in the plant, while active resistance relies on defenses that
are induced after infection or attack.
11

Characteristics of induced systemic resistance
• The defensive capacity of the plant is enhanced through microbial
stimulations or similar stresses.
• Systemic change in physiology and gene expression.
• The enhanced defensive capacity is expressed systemically
throughout the plant.
• Triggered by nonpathogenic agents.
• Involves jasmonic acid, ethylene accumulation.
• Induced systemic resistance is active against fungi, bacteria,
viruses and, sometimes, nematodes and insects.
• Local or Systemic protection.
12

• Enhancement of activities rather than elimination of the
pathogens.
• Once induced systemic resistance is maintained for prolonged
periods.
• Resistance against broad spectrum of pathogens.
• Time dependant i.e. resistance is established only after certain
metabolic changes occur in the host during a specific interval
following the inducing inoculation.
• Rapidity and intensity depend upon the concentration of inducers
and the number of host cells that are affected.
L. C. van Loon, (2007). Eur J Plant Pathol 119:243–254.
13

Fig.2. Biologically induced resistance triggered by pathogen infection (red arrow),
insect herbivory (blue arrow), and beneficial microbes (purple arrows). Here,
Secondary pathogen infections (2˚) of induced plant tissues cause significantly less
damage than those primary (1˚) infected or infested tissues.
Primary infection
secondary infection
Pieterse et al. (2014). The Annual Review of Phytopathology, 52:347–75 14

Fig.3.Signal transduction pathways leading to pathogen-induced systemic acquired resistance
(SAR) and Rhizo bacteria-mediated induced systemic resistance (ISR) in Arabidopsis thaliana.
Anelise Beneduzi et al., (2012).Genetics and Molecular Biology, 35, 4 (suppl), 1044-1051. 15
Mechanisms of induced systemic resistance

NPR1 : Non-expresser of PR genes
 NPR1 is key & positive regulator of ISR.
 Expression of NPR1 is induced by pathogen infection or
treatment with defense-inducing compounds.
 NPR1 mutants are susceptible to various pathogens.
 The non-expresser of PR genes (NPR1) has emerged as a
good candidate to provide broad-spectrum resistance.
 NPR1 is a regulatory protein that activates & expression
of PR genes.
 It participates in the jasmonate and ethylene regulation,
SA- independent induced systemic resistance (ISR).
16

Fig.4. Factors affecting the expression of induced resistance in practice
Dale R. Walters et al. (2013), Journal of Experimental Botany, Vol. 64: 1263–1280. 17

Passive resistance Active resistance
Physical
barriers
Chemical
barriers
Rapid
defense Delayed
defense
 Structure of
epidermis
 Structure of
stomata
 Mechanical
tissues
 Phyto anticipins
 pH  Membrane permeability loss
 Oxidative burst
 Fortification cell wall
 HR
 Phytoalexin – production
 Systemic Acquired
Resistance (SAR)
 Induced Systemic
Resistance (ISR)
Dube, 2016
Disease Resistance
Types of Induced Resistance
18

Fig.5. A pictorial comparison of the two best characterized forms of induced
resistance in plants, Systemic acquired resistance and Induced systemic
resistance. Both which lead to similar phenotypic responses.
Vallad and Goodman (2004). Crop Science, 44:1920–1934. 19

Alpha- Linolenic
acid
peroxide
allene oxide
12-oxophytodienoic acid (OPDA)
Jasmonic acid
Fig.6. Jasmonic Acid (JA) Biosynthesis Pathway
In
Plastid
(chloroplast)
http://youtube/03cduXJddnM
Oxygeneted by
Lipoxygenase
Rearrangement by Allene
oxide cyclase (AOC)
Cyclize in presence of
Allene oxide synthase
20

L-Methionine S- adenosyl methionine (SAM)
1-aminocyclopropane-
1-carboxylate (ACC)
Ethylene
(2HC=CH2)
Fig.7. Ethylene (ET) Biosynthesis pathway
http://www.aribidopsis.com/2016/05/Ethylene-biosynthesis-pathway-ch03
AdoMet synthatase
ACC synthase
ACC oxydase
ATP
CO2 + HCN
PPi + Pi
1/2
O2
21

Role of Siderophores in induce systemic resistance (ISR)
Siderophores are low-molecular-weight molecules that
are secreted by microorganisms to take up iron from the
environment.
Their modes of action in suppression of disease were
thought to be solely based on competition for iron with the
pathogen.
Interestingly, siderophores can induce systemic resistance
(ISR).
One may therefore consider whether the mode of action
of other bacterial metabolites that have been implicated in
disease suppression also involves triggering of systemic
resistance mediated by rhizobacteria.
P.A.H.M. Bakker et al.,(2003),Can. J. Plant Patho. Vol. 25. 22

Plant growth-promoting rhizobacteria elicited ISR
Over the past decade, specific strains of plant growth
promoting rhizobacteria (PGPR) have been shown to induce
systemic resistance against a broad spectrum of pathogens such
as fungi, bacteria and viruses PGPR-elicited induced systemic
resistance (ISR).
ISR elicited by PGPR render non infected parts of
previously induced plants more resistant to infection by
pathogens and are effective against a broad spectrum of root and
foliar pathogens.
Shouan Zhang et al., (2002). Biological Control, 25 :288–296.
23

Fig.8. Plant growth–promoting effect of plant growth–promoting rhizobacteria (PGPR) strain
Pseudomonas fluorescens WCS417r on Arabidopsis. (a) Colonization of Arabidopsis roots by P.
fluorescens WCS417r increases shoot biomass and stimulates lateral root formation and root hair
development. (b) P. fluorescens WCS417r–induced changes in root architecture are stimulated
via auxin-dependent responses in the Arabidopsis root.
•Pieterse et al. (2014). The Annual Review of Phytopathology, 52:347–75.
•The Annual Review of Phytopathology is online at phyto.annualreviews.org 24

Fig.9. Working model explaining the possible involvement of JA and ethylene in P.
fluorescens WCS417r-mediated ISR in Arabidopsis.
Pieterse et al. (2001) European Journal of Plant Pathology, 107: 51-61 25

Mechanisms of plant growth promotion by rhizobacteria
• Nitrogen fixation
• Ion uptake:
Iron, zinc, other essential micronutrients, Phosphate
• Production of plant hormones:
Auxins, gibberellins, cytokinins, ethylene
• Modulation of plant development
• ACC deaminase (1-aminocyclopropane-1-carboxylic
acid (ACC)).
• ‘Elicitors’.
26
Van loon et al. (2007) Eur J Plant Pathol 119:243–254.

Table: 01 Rhizobacteria-mediated induced systemic resistance in plant
species investigated
Van loon et al. (1998).
Netherlands. 27

Van loon et al. (1998).
Netherlands. 28

Table: 02 Differential induction of systemic
resistance by Pseudomonas spp. strains
Netherlands. Van loon et al. (1998).
29

Table: 03 Bacterial determinants of induced systemic resistance by fluorescent
Pseudomonas spp. in different host plants Determinant Pseudomonas spp. strain
Peter A. H. M. Bakker et al.,(2007). 30
The Netherlands.

Fig.10. Model for Trichoderma-induced resistance (TISR) against Botrytis cinerea in
tomato. Root colonization with Trichoderma primes leaf tissues for enhanced activation
of JA-regulated defense responses leading to a higher resistance to the necrotroph.
Intact JA, SA, and ABA signaling pathways are required for TISR development.
Ainhoa Martínez-Medina et al., (2013), Frontier in plant science, 4 : 1-12. 31

Fig.11. Diagram of the main phases involved in root colonization by beneficial
soil borne bacteria and their functions.
•Pieterse et al. (2014). The Annual Review of Phytopathology, 52:347–75.
•The Annual Review of Phytopathology is online at phyto.annualreviews.org 32

• (I) Plant roots selectively secrete organic compounds that
function as semiochemicals for the assembly of the root
microbiome. Selected bacterial strains from the bulk soil
communities specifically respond to host signals and reprogram
to express traits related to root colonization.
• Microbes that have evolved as endophytes commonly enter the
root interior through cracks in the root epidermis or through root
hairs (inset).
• In phase I, local immune responses in host roots are transiently
suppressed by epiphytic or endophytic plant growth–promoting
rhizobacteria (PGPR), allowing bacteria to propagate on the root
epidermis or intracellularly.
THE ROOTS OF INDUCED SYSTEMIC RESISTANCE: EARLY
SIGNALING EVENT
33

• II) Once PGPR are established on the root, cell wall
polysaccharides from the host function as environmental cues to
promote biofilm formation on the root surface. Within the biofilm
matrix, individual members and/or microbial consortia integrate
host and self-derived signals to activate processes in the plant that
lead to enhanced plant growth and induced systemic resistance
(ISR).
• In addition, root microbiota protect root tissues against soil borne
pathogens via the production of antibiotics and competition for
nutrients and niches.
• (III) Early root responses to beneficial microbes are locally
expressed in the epidermis and are subsequently communicated to
the inner cell layers and to the aboveground plant parts via yet
elusive long-distance molecules, where these signals confer ISR.
34

Aswathi et al.(2019)
01.
Tamilnadu, India. 35

Table 04: Effect of Bacillus subtilis strain VB1 (talc formulation) on the incidence of
wilt under glass house conditions
Sr. No Treatment Wilt incidence
(%) 60 DAS
01 ST with Bacillus subtilis VB1 @ 10g/kg of seeds 22.32bcd
02 SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 24.43bc
03 ST (10g/kg of seeds) + SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 20.39cd
04 ST (10g/kg of seeds) + SA with Bacillus subtilis VB1(Basal & top dressing) @
2.5kg/ha
16.33e
05 ST with Pseudomonas fluorescens Pf1 @ 10g/kg of seeds 23.87bc
06 SA with Pseudomonas fluorescens Pf1 (Basal) @ 2.5kg/ha 26.46b
07 ST (10g/kg of seeds) +SA with Pseudomonas fluorescens Pf1 (Basal) @2.5kg/ha 20.57cd
08 ST (10g/kg of seeds) + SA with Pseudomonas fluorescens Pf1 (Basal & top
dressing) @ 2.5kg/ha
19.32de
09 ST with Carbendazim @ 2g/kg of seeds 22.55bcd
10 ST (2g/kg of seeds) + SD with Carbendazim @ 0.1% 20.34cd
11 Untreated pathogen inoculated control 35.56a
ST – Seed treatment SA – Soil application SD – Soil drenching Values are mean of three replications
Means followed by a common letter are not significantly different at 5 % level by DMRT 36

Table 05: Induction of Peroxidase (PO) activity in coriander plants applied with
biocontrol agents under glass house conditions
Sr.
No
Treatment Change in absorbance at 420 nm/min/g of sample
01 ST with Bacillus subtilis VB1 @ 10g/kg of seeds 0.687d 0891bc 1.136f 1.521e 1.310d
02 SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 0.783c 0.972b 1.239e 1.257g 1.10e
03 ST (10g/kg of seeds) + SA with Bacillus subtilis VB1 (Basal)
@ 2.5kg/ha
0.987a 1.133a 1.843b 1.930b 1.725b
04 ST (10g/kg of seeds) + SA with Bacillus subtilis VB1(Basal &
top dressing) @ 2.5kg/ha
0.892b 1.153a 1.991a 2.175a 1.967a
05 ST with Pseudomonas fluorescens Pf1 @ 10g/kg of seeds 0.582e 0.765d 0.996g 1.296g 0.918f
06 SA with Pseudomonas fluorescens Pf1 (Basal) @ 2.5kg/ha 0.334f 0.658e 1.099f 1.375f 1.017e
07 ST (10g/kg of seeds) +SA with Pseudomonas fluorescens Pf1
(Basal) @2.5kg/ha
0.967a 1.142a 1.675c 1.943b 1.586c
08 ST (10g/kg of seeds) + SA with Pseudomonas fluorescens Pf1
(Basal & top dressing) @ 2.5kg/ha
0.980a 1.101a 1.675c 2.031a 1.930a
09 ST with Carbendazim @ 2g/kg of seeds 0.279f 0.751d 1.491d 1.875c 1.567c
10 ST (2g/kg of seeds) + SD with Carbendazim @ 0.1% 0.582e 0.965b 1.296e 1.596e 1.118e
11 Untreated pathogen inoculated control 0.134g 0.828c 1.399e 1.775d 1.217d
0 DAS 3 DAS 5 DAS 7 DAS 9 DAS
ST – Seed treatment SA – Soil application SD – Soil drenching Values are mean of three replications Means
followed by a common letter are not significantly different at 5 % level by DMRT 37

Table 06: Induction of Poly peroxidase (PPO) activity in coriander plants applied with
Sr.
No
01 ST with Bacillus subtilis VB1 @ 10g/kg of seeds 0.884 1.461d 1.960c 2.256e 1.944e
02 SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 0.804e 1.361e 1.776d 2.017f 1.762f
@ 2.5kg/ha
0.952d 1.791b 2.323b 2.503c 2.445b
1.246a 1.893a 2.456a 2.830a 2.509a
05 ST with Pseudomonas fluorescens Pf1 @ 10g/kg of seeds 0.834e 1.321e 1.843d 2.031f 2.376c
06 SA with Pseudomonas fluorescens Pf1 (Basal) @ 2.5kg/ha 0.932d 1.454d 1.932c 2.385d 2.068d
(Basal) @2.5kg/ha
1.043c 1.543c 1.864d 2.458d 2.142d
1.154b 1.743b 2.244b 2.643b 2.245d
09 ST with Carbendazim @ 2g/kg of seeds 0.787f 1.100f 1.572e 2.044f 1.702f
10 ST (2g/kg of seeds) + SD with Carbendazim @ 0.1% 0.792ef 1.021g 1.402f 1.672g 1.503g
11 Untreated pathogen inoculated control 0.691g 0.931h 1.281g 1.540h 1.401h

Table 07: Induction of Phenyl alanine ammonia lyase (PAL) activity in coriander plants applied with
Sr.
No
01 ST with Bacillus subtilis VB1 @ 10g/kg of seeds 0.742d 0.847f 0.993fg 1.356g 1.023e
02 SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 0.695e 0.783fg 0.902g 0.957h 0.873f
@ 2.5kg/ha
0.796d 1.320c 2.033a 2.205c 2.092b
04 ST (10g/kg of seeds) + SA with Bacillus subtilis VB1(Basal
& top dressing) @ 2.5kg/ha
1.036a 1.654a 1.929ab 2.75a 2.534a
05 ST with Pseudomonas fluorescens Pf1 @ 10g/kg of seeds 0.723de 0.884f 1.032f 1.442f 1.032e
06 SA with Pseudomonas fluorescens Pf1 (Basal) @ 2.5kg/ha 0.748d 1.088e 1.487e 1.754e 1.533d
(Basal) @2.5kg/ha
0.801c 1.139d 1.638d 2.023d 1.836c
08 ST (10g/kg of seeds) + SA with Pseudomonas fluorescens
Pf1 (Basal & top dressing) @ 2.5kg/ha
0.988b 1.454b 1.856c 2.476b 2.043b
09 ST with Carbendazim @ 2g/kg of seeds 0.689e 0.774g 0.867h 0.946h 0.845f
10 ST (2g/kg of seeds) + SD with Carbendazim @ 0.1% 0.698e 0.786fg 0.799i 0.841i 0.765g
11 Untreated pathogen inoculated control 0.667f 0.789fg 0.791i 0.895h 0.691h

Table 08: Induction of f Phenol content in coriander plants applied with biocontrol agents under glass house
conditions
Sr.
No
Treatment Phenol content (mg/g of sample)
01 ST with Bacillus subtilis VB1 @ 10g/kg of seeds 4.274f 4.876d 5.531c 5.839d 5.231f
02 SA with Bacillus subtilis VB1 (Basal) @ 2.5kg/ha 4.514c 4.893d 5.439d 5.717e 5.482d
@ 2.5kg/ha
4.393e 4.984c 5.432d 5.849d 5.029g
4.712a 5.289b 5.832a 6.396a 5.99a
05 ST with Pseudomonas fluorescens Pf1 @ 10g/kg of seeds 4.309e 4.790e 5.056f 5.467f 5.065g
06 SA with Pseudomonas fluorescens Pf1 (Basal) @ 2.5kg/ha 4.401d 4.788e 5.478d 5.976c 5.687b
(Basal) @2.5kg/ha
4.603b 4.988c 5.487d 5.978c 5.530c
4.754a 5.580a 5.854a 6.238b 5.965a
09 ST with Carbendazim @ 2g/kg of seeds 4.384e 4.734e 5.642b 5.893d 5.391e
10 ST (2g/kg of seeds) + SD with Carbendazim @ 0.1% 4.390e 4.832d 5.358e 5.958c 4.732i
11 Untreated pathogen inoculated control 4.274f 4.637f 5.028f 5.268g 4.890h

Madhavi et al.(2018)
02.
41
Andhra Pradesh, India.

Treatment PO activity (change in absorbance
(OD/min/g)
PAL activity(change in absorbance
(OD/min/g)
PDI
Days after inoculation Days after inoculation
0 1 2 3 4 0 1 2 3 4
T1- Seed treatment with
P. fluorescens @10ml-1kg
1.17 1.27 1.43 1.43 1.37 0.31 0.47 0.63 0.47 0.43 68.0
T2- Seed treatment with
T. harzianum @10ml-1kg
1.07 1.17 1.27 1.23 1.27 0.31 0.43 0.47 0.40 0.31 67.3
T3- Seed treatment + foliar
application of P. fluorescens @
5ml-1
1.17 1.47 1.83 1.87 2.00 0.31 1.07 1.13 1.17 1.22 44.7
application of T. harzianum @
5ml-1
1.13 1.33 1.63 1.77 1.93 0.31 0.93 0.97 0.99 1.07 51.7
T5- Seed treatment P.
fluorescens@ 5ml-1kg +
T. harzianum @ 5ml-1
1.10 1.93 2.13 2.20 2.27 0.31 1.23 1.23 1.27 1.29 62.7
application of P. fluorescens @
5ml-1 + T. harzianum @ 5ml-1
1.13 2.17 2.43 2.5 7 2.73 0.31 1.43 1.63 1.68 1.72 37.7
T7- Control 1.10 1.17 1.07 1.07 1.23 0.31 0.31 0.32 0.31 0.31 89.0
SEm± 0.03 0.03 0.03 0.03 0.03 0.00 0.02 0.02 0.02 0.02 0.17
CD NS 0.09 0.09 0.11 0.11 NS 0.06 0.08 0.09 0.05 5.34
Table 09: Changes in peroxidase (PO) and phenylalanine ammonia lyase (PAL) activity in maize leaf sheaths due to seed
treatment with Pseudomonas fluorescens and Trichoderma harzianum alone and in combination, challenge inoculated with
Rhizoctonia solani f.sp. sasakii
42

Rajkumar et al.(2018)
03.
43
Raichur, India

Table:10 Induction of peroxidase activity in chilli by B. subtilis (BS) isolates
challenge inoculated with F. solani
Sr.
No
Treatment Change in absorbance at 470 nm/min/mg of protein
01 BS5+FS - - 0.14 0.45 0.81 0.73
02 BS7+FS - - 0.26 0.59 0.89 0.81
03 BS9+FS - - 0.22 0.49 0.86 0.79
04 BS16+FS - - 0.23 0.69 1.04 0.98
05 BS30+FS - - 0.24 0.62 0.99 0.93
06 Inoculated (FS) - - 0.14 0.43 0.76 0.69
07 Un inoculated - - 0.12 0.39 0.65 0.60
08 S.Em ± - - 0.01 0.01 0.01 0.01
09 C.D at 1 % - - 0.02 0.03 0.05 0.04
0 DAS 1 DAS 3 DAS 5 DAS 7 DAS 9 DAS
44

Table:11 Induction of polyphenol oxidase activity in chilli by B. subtilis (BS)
isolates challenge inoculated with F. solani (FS)
Sr.
No
Treatment Change in absorbance at 470 nm/min/mg of protein
01 BS5+FS - - 0.08 0.38 0.48 0.41
02 BS7+FS - - 0.16 0.40 0.71 0.65
03 BS9+FS - - 0.10 0.39 0.56 0.51
04 BS16+FS - - 0.18 0.51 0.89 0.79
05 BS30+FS - - 0.13 0.49 0.79 0.71
06 Inoculated (FS) - - 0.06 0.34 0.45 0.40
07 Un inoculated - - 0.05 0.20 0.40 0.34
08 S.Em ± - - 0.01 0.01 0.01 0.01
09 C.D at 1 % - - 0.04 0.03 0.03 0.02
45

Table:12 Induction of phenylalanine ammonia lyase activity in chilli by B. subtilis
(BS) isolates challenge inoculated with F. solani (FS)
Sr.
No
Treatment nmol trans- cinamic acid /hr /mg protein
01 BS5+FS - - 14.20 22.80 71.80 65.77
02 BS7+FS - - 14.67 20.57 78.47 74.13
03 BS9+FS - - 13.84 21.57 74.47 67.67
04 BS16+FS - - 16.67 28.83 80.45 78.53
05 BS30+FS - - 15.27 26.60 78.50 74.71
06 Inoculated (FS) - - 11.30 19.97 62.43 60.29
07 Un inoculated - - 9.20 16.63 50.42 47.60
08 S.Em ± - - 0.05 0.13 0.22 0.37
09 C.D at 1 % - - 0.21 0.57 0.97 1.58
46

Gujarat, India.
04.
47
Patel and Saraf (2017)

Table.13. Fusarium oxysporum f. sp. lycopersici incidence on tomato
plant.
Treatment Disease incidence (%)
Control -
FOL 83 ± 0.68
MSST -
MSST + FOL 12.4 ± 0.64
LSD 11.9
48
All the values represent the mean of three replicates ± standard deviation.
different letters denote a statistically significant difference according to
Duncan's Multiple range test; least significant difference (lsd) at p ≤ 0.05
FOL= Fusarium oxysporum f. sp. Lycopersici
MSST=Trichoderma asperellum strain

Figure 12. Po and PPo activity induced by Trichoderma asperellum
Msst in tomato plant treated with or without F. oxysporum f. sp.
lycopersici
49

50
Figure 13. PAL activity induced by Trichoderma asperellum Msst
in tomato plant treated with or without F. oxysporum f. sp.
lycopersici

Tehran, Iran
05.
51
Fotoohiyan et al. (2015)

Fig.14. Effect of the treatments of Trichoderma harzianum isolates and
Verticillium dahliae on wilt disease reduction in pistachio seedlings under
greenhouse conditions one month after inoculation. V= V . dahliae and Tr
= T . harzianum
52

53
Verticillium dahliae PO Activities in pistachio seedlings under
greenhouse conditions one month after inoculation. V= V . dahliae and Tr
= T . harzianum

54
Verticillium dahliae PAL Activities in pistachio seedlings under greenhouse
conditions one month after inoculation. V= V . dahliae and Tr = T . harzianum

55
Verticillium dahliae Total phenolic content in pistachio seedlings under
greenhouse conditions one month after inoculation. V= V . dahliae and Tr = T .
harzianum

Giza, Egypt.
06.
56
Monaim et al. (2015)

Table.14: Effect of fodder beet seeds treatment with potassium salts on damping off, root rot/wilt diseases during 2013/14 and 2014/15 growing seasons
under field conditions.
Potassium salts Concen. (g/L)
Season 2013-14 Season 2014-15
% Damping off % Root rot/ wilt % Damping off % Root rot/ wilt
K2HPO4 5 15.24 15.24 12.35 12.24
10 12.35 10.32 10.33 8.25
20 10.33 7.36 8.21 6.36
Mean 12.64 10.97 10.30 8.95
KHCO3 5 25.36 19.35 24.14 18.52
10 20.55 15.34 18.25 12.36
20 14.86 16.25 13.24 14.96
Mean 20.26 16.98 18.54 15.28
K2SO4 5 20.14 18.69 17.67 17.25
10 14.25 10.24 10.25 9.58
20 16.36 12.36 13.24 12.36
Mean 16.92 13.76 13.72 13.06
K2SiO3 5 10.25 12.36 8.56 10.25
10 7.36 6.36 6.25 5.36
20 5.28 5.45 5.28 5.56
Mean 7.63 8.06 6.70 7.06
Control 35.26 25.36 30.25 26.35
LSD at 0.05 for:
Potassium salts (A)= 2.65 2.44 2.47 2.31
Concentrations (B)= 3.01 2.09 2.85 2.59
Pathogenic fungi (C)= 2.69 2.14 2.54 2.51
Interaction (A×B×C)= 7.48 6.51 7.01 6.78

Figure.18: Effect of potassium salts on peroxidase activity (PO) in
inoculated fodder beet plants. Mean ± SDs for nine plants per
treatment are shown. Different letters indicate significant differences
between treatments according to LSD test (P ≤ 0.05).
58

Figure.19: Effect of potassium salts on polyphenol oxidase activity
(PPO) in inoculated fodder beet plants. Mean ± SDs for nine plants per
59

Figure.20: Effect of potassium salts on phenylalanine ammonia lyase
activity (PAL) in inoculated fodder beet plants. Mean ± SDs for nine
plants per treatment are shown. Different letters indicate significant
differences between treatments according to LSD test (P ≤ 0.05).
60

Figure.21: Effect of potassium salts on tyrosine ammonia lyase activity
(TAL) in inoculated fodder beet plants. Mean ± SDs for nine plants per
61

Figure.22: Effect of potassium salts on total phenol content (TPC) in
inoculated fodder beet plants. Mean ± SDs for nine plants per
62

Banglore, India.
07.
63
Sivakumar et al. (2013)

Treatments Peroxidase (PO) activity (changes in absorbance/min/g of
tissue)
Days after inoculation (DAI)
2 4 6 8 10 12
Root dipping of
B. megaterium
1.41 1.51 2.01 1.72 1.61 1.55
Root dipping of
B. megaterium +
R. solanacearum
1.72 1.82 2.75 2.26 2.11 2.01
R. solanacearum 1.12 1.14 1.62 1.53 1.52 1.34
Control 0.54 0.72 0.75 1.03 1.15 1.21
64
Table.15 Peroxidase activity in brinjal plants treated with Bacillus
megaterium

Treatments Polyphenol oxidase (PPO) activity (changes in
absorbance/min/g of tissue)
2 4 6 8 10 12
Root dipping of
B. megaterium
0.32 0.44 0.77 0.70 0.49 0.43
Root dipping of
B. megaterium +
R. solanacearum
0.68 0.70 0.91 0.59 0.57 0.50
R. solanacearum 0.25 0.39 0.52 0.31 0.25 0.15
Control 0.10 0.11 0.13 0.14 0.16 0.13
65
Table.16. Polyphenol oxidase activity in brinjal plants treated
with Bacillus megaterium

Treatments Phenol content µg g–1 of plant tissue
2 4 6 8 10 12
Root dipping of
B. megaterium
145 146 155 149 148 147
Root dipping of
B. megaterium +
R. solanacearum
149 150 173 164 153 154
R. solanacearum 142 143 149 114 143 142
Control 120 122 121 123 123 122
66
Table.17. Phenol content in brinjal plants treated with Bacillus
megaterium

08.
Seleim et al. (2011)
Assiut, Egypt. 67

Fig.23. Disease reduction percentage of treated tomato plants with
PGPR strains under greenhouse conditions.
68
Disease
reduction
percentage
%
80%
68% 60%
8%

Table.18. The average disease incidence and disease reduction percentage of treated tomato plant with PGPR
strains.
Treatment
PGPR Concentration Disease incidence % Disease reduction %
Pseudomonas putida
108 45FG 52.63
107 45FG 52.63
106 50EFG 47.37
105 65BCDE 31.58
Pseudomonas fluorescens
108 40G 57.90
10b 40G 57.90
106 45FG 52.63
105 45FG 52.63
Bacillus subtilis
108 45FG 52.63
107 60CDEF 36.84
106 60CDEF 36.84
105 70BCD 26.32
Enterobacter aerogenes
108 50EFG 47.37
107 75BC 21.10
106 80AB 15.79
105 45FG 15.79
Mixture if PGPR strains
108 80AB 52.63
107 55DEFG 42.11
106 50EFG 47.37
105 65BCDE 31.58
Infected control 95A
69

09.
Saravanakumar et al. (2007)
Coimbatore, India. 70

Table.19. Effect of foliar application of PGPR bioformulations on disease index of blister blight in tea plants.
Treatment 0 DAS 15 DAS 30 DAS 45 DAS 60 DAS 75 DAS 90 DAS 105 DAS Mean
disease
index(%)
Pf1 at 7
DI
5.86b
(13.87)
17.6g
(24.75)
29.62f
(32.96)
15.54g
(23.18)
16.63f
(26.04)
13.58f
(21.58)
14.13f
(22.00)
15.66f
(23.28)
16.06f
Pf1 at 14
DI
6.02a
(14.07)
28.05e
(31.95)
37.84d
(37.95)
22.26d
(27.13)
23.10d
(27.32)
18.86d
(25.71)
21.05d
(27.27)
24.22d
(29.46)
23.15d
Pf1 at 21
DI
5.98b
(14.02)
36.54d
(37.18)
46.50c
(42.99)
26.64b
(30.48)
25.25c
(30.15)
21.67c
(27.72)
25.06c
(30.32)
26.66c
(31.07)
26.89c
B. Subtilis
at 7 DI
6.52a
(14.68)
24.23f
(29.45)
33.18e
(35.16)
18.00e
(25.08)
20.66e
(27.01)
17.32d
(24.57)
21.02d
(27.25)
21.22e
(27.41)
20.25e
B. Subtilis
at 14 DI
5.46b
(13.36)
37.84c
(37.95)
49.91b
(44.94)
24.63c
(29.74)
25.88c
(30.56)
21.00c
(27.25)
24.65c
(29.74)
25.99c
(30.63)
26.90c
B.
Subtilis at
21 DI
5.78b
(13.77)
39.49b
(38.92)
49.03b
(47.32)
28.14b
(32.02)
27.22b
(31.43)
26.10b
(30.71)
27.73b
(31.75)
36.66b
(37.25)
30.00b
Hexacona
zole(0.25)
6.25ª
(14.35)
14.85h
(22.29)
29.10f
(32.63)
17.94f
(25.03)
19.20e
(25.96)
15.9e
(23.48)
16.42e
(23.84)
16.08f
(23.61)
16.90f
Control 5.83b
(13.83)
42.24ª
(40.53)
79.00a
(62.74)
49.33ª
(44.61)
39.54ª
(38.95)
34.90a
(36.20)
40.80a
(39.69)
45.25a
(42.27)
42.09a
71

10.
Zhinong Yan et al. (2002)
Auburn, Alabama. 72

Fig.24. Effect of selected plant growth-promoting rhizobacteria (PGPR) strains
on tomato late blight control under greenhouse conditions.. Disease was visually
measured by percent leaf area covered with late blight lesions. Data are the
means of three experiments. Different letters indicate significant differences
among treatments according to a least significant difference test (P = 0.05).
73

Fig.25. Percent germination of sporangia of Phytophthora infestans on tomato
leaves induced with plant growth-promoting rhizobacteria (PGPR) strains SE34
and 89B61, β-amino butyric acid (BABA), and pathogen. Data are means of two
experiments. Different letters indicate significant differences among treatments
according to a least significant difference test (P = 0.05).
74

Fig.26. Zoospore germination of Phytophthora infestans on tomato leaves
induced with plant growth-promoting rhizobacteria PGPR strains SE34 and
89B61, β-amino butyric acid (BABA), and pathogen. Data are means of two
experiments. Different letters indicate significant differences among treatments
according to a least significant difference test (P = 0.05).
75

Induced Systemic Resistance is an essential component of plant
defense mechanism.
Among various biocontrol agents, Pseudomonas fluorescens,
Trichoderma sp., Bacillus sp. has a special importance in plant defense
mechanism by ISR through production of different defense enzymes like
PO, PPO, PAL, with JA/EA pathway.
They helps in minimizing disease caused by pathogens like
Fusarium sp., Rhizoctonia solani, Colletotrichum sp., Verticillium dahliae,
Ralstonia solanacearum, Pythium aphanidermatum, Phytophthora
infestans etc. Some Potassium salts also inducing resistance against
damping off and wilt diseases and in some case studies combination of
PGPR strains along with the pathogens also playing the role in inducing
defense mechanism.
ISR through application of Pseudomonas fluorescens, Trichoderma
sp., Bacillus sp. will be employed for efficient and eco-friendly disease
management and enhance profitability to farming community.
Conclusion
76

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