compatibility and Tolerance in Trichoderma spp. and Pseudomonas fluorescens with fungicides

2. SEMINAR ON
Fungicides Tolerance in Trichoderma spp. and Pseudomonas fluorescens
Pansuriya Bhavin J.
M.Sc. (Agri.) Plant Pathology,
C. P. College of Agriculture., S.D.A.U.,
Sardarkrushinagar.
Reg. No.:04-AGRMA-02244-2020
S P E A K E R
Dr. N. R. Patel
Associate Research Scientist
( Pl. Path.),
Seed Spices Research Station.
S.D.A.U., Jagudan.
M A J O R A D V I S O R
Dr. P. S. Patel
Associate Professor,
Department of Entomology,
C. P. College of Agriculture,
S.D.A.U., Sardarkrushinagar.
M I N O R A D V I S O R

3. SEMINAR OUTLINE
1
Bio Control Agents
Drawback of bio agents & its
solution
3
2
5
4
Conclusion
Introduction
Case studies

4. Introduction
 The Green Revolution in India (1960) refers to a period when
Indian agriculture was converted into an industrial system due to
the adoption of modern methods and technology such as the use
of high yielding variety (HYV) seeds, irrigation facilities,
pesticides and fertilizers.
 Generally, hybrid and high yielding varieties are more susceptible
to disease than wild varieties.
 In order to protect crops from different types of diseases caused
by microorganisms, the farmers used pesticides at a high amount.
1

5.  The widespread use of pesticides has resulted in resurgence
of pests, environmental pollution, development of resistant
strains, etc.
 The usage of a high quantity of fungicides and insecticides
incorporated toxicity in the plants.
 Eventually, The benefits of the Green Revolution have been
coupled with unanticipated harmful consequences from
chemical pollution.
2

6.  Pesticides are necessary at present but are not a long term
solution to crop, human and animal health.
 Besides their non-target effects and hazardous nature, they are
becoming more expensive and some are loosing their
effectiveness because of development of resistant strains.
 According to WHO estimate (2012) approximately one million
people are taken ill every year with pesticide poisoning and up
to 20,000 of those die in agony.
Source : SNApplied Sciences, 2019, 1:1446
Evils of Pesticides
3

7.  Four decades after Indian farmers began increasing production
using pesticides and fertilizers, they are starting to have second
thoughts about the change.
 In 2008, Researchers at Punjab University discovered DNA
damage in 30 percent of Indian farmers who treated plants with
herbicides and pesticides.
 An additional study found heavy metals and pesticide chemicals in
all products including drinking water. These substances are
harmful and can cause serious health problems.
 Some of these problems may occur because some farmers may not
know how to handle and dispose of toxic chemicals. They may
also harm the environment by using too many of those products.
Source : Lee, Kevin(2018) Harmful Effects of the Green Revolution" @ sciencing.com 4

8.  Due to hazardous effects of chemical pesticides, biological
management of disease is getting more important than
chemical methods.
 Bio-agents reduce excessive use and misuse of synthetic
fungicide and development of fungicide resistance in pathogens
(Kumar et al., 2017).
 Currently several bio control agents are recognized and available
as following : Trichoderma, Gliocladium and Ampelomyces
(fungal bio agents) & Pseudomonas, Bacillus, Agrobacterinum
(Bacterial bio agents).
 Among them Trichoderma & Pseudomonas spp. are most
commonly used as widely available in market.
5

9. Trichoderma spp.
It is free living, Ubiquitous, Non polluting, Non
phytotoxic, easily accessible and highly
proliferating fungi.
Trichoderma spp. are abundantly present in the
soil, rhizosphere and are mycoparasitic of several
soil-borne plant pathogens.
Several species of Trichoderma produce volatile
and non-volatile antibiotics and enzymes which are
antagonistic to phytopathogenic fungi and
nematodes.
6

10.  The fungus is effective against pathogens causing
various diseases of the root region of plants viz., collar
rot, foot rot, damping off etc.
 In the rhizosphere, some strains of Trichoderma spp.
release metabolites which improve the growth of
seedling and it also causes resistance against abiotic
stress.
 Trichoderma spp. have great potential against soil-
borne pathogens and it may be able to replace
chemical pesticides in near future (Manish and
Shabbir, 2017).
7

11. Pseudomonas fluorescens
 Pseudomonas fluorescens are important group
of bacteria that play major role in the induced
systemic resistance, biological control of plant
pathogens and plant growth promotion etc.
 It encompasses a group of common,
nonpathogenic saprophytes that colonize soil,
water and plant surface environments.
 It is rod shaped bacteria have multiple polar
flagella.
8

12.  Growth rapidly in vitro and to be mass produced.
 Rapidly utilize seed and root exudates and colonizes
and multiplies in the rhizosphere and spermosphere
environment and in the interior of the plant .
 Produce a wide spectrum of bioactive metabolites.
 Compete aggressively with other microorganisms.
 Adapt to environmental stresses.
Pseudomonas fluorescens as bio control agent
Electron Microscopic view
9

13. Induced systemic resistance of the host plant
Plant Growth promotion
Competition
Antibiosis
Mycoparasitism
Mode of action
10

14. Is it long term solution?
11

15. Depend on
environment conditions
Draw back of
bio agents
Lack of availability and
awareness
Used against specific
diseases
Slow and Less effective
than chemicals
Short shelf life
Only a preventive
measure 12

16.  Bio control agents can not manage the disease completely when
large scale infection is already established in the field, farmers
favoured fungicides for managing the crop diseases.
 Indiscriminate use of pesticides has resulted in accumulation of toxic
compounds which affects adversely to humans health and
environment.
 Only application of bio-agent is not the answer for management of
plant diseases.
 From the above conclusion, only chemicals or only bio-agents can
not give long term solution of plant disease management.
13

17.  Integration of bio-agents with fungicides may enhance the
effectiveness of disease control and provide better management of
soil borne diseases (Papavizas and Lewis, 1981).
 To develop an effective disease management programme,
knowledge about the compatibility of potential bio-agents with
commonly used agrochemicals are essential.
 As fungicides should have inhibitory effect on the pathogen but
should not have deleterious effect on the antagonists.
 An understanding of the effect of fungicides on the pathogen and
the antagonists would provide information for the selection of
fungicides through compatibility studies in vitro.
14
So, What is the solution?

18.  Combining antagonists with synthetic chemicals eliminates the
chance of resistance development and reduces the fungicide
application ( Wedajo, 2015).
 In several disease management strategies, the addition of
fungicide at reduced rates in combination with bio-control
agents has significantly enhanced disease control, compared to
treatments with only bio-control agents (Kumar et al., 2017).
 In addition, this strategy may display even better control of
resistant strains of fungal pathogens and may help the
commercial growers to reduce the amount of fungicide use, thus
lowering the amount of chemical residue in the marketed
products (Nandini et al., 2018).
15

19. Benefits of compatibility
Eliminates the chance of
resistance development
Reduce cost of the
formulations
Can control more than
one disease at same time
Enhance the effectiveness
of disease control program
Provide better disease
suppression of pathogen as
compare to sole fungicides
Reduce the amount of fungicide
use and chemical residue
16

20. Case studies

21. Fungicides tolerance in Trichoderma spp. in vitro

22. Table 1: Effect of fungicides on mycelial growth of Trichoderma asperellum
Kerala Kumar et al., (2017)
18
Sr. No. Fungicides
Mycelial inhibition (%) at
100 ppm 200 ppm 400 ppm 800 ppm
1 Metalaxyl M-8% + Mancozeb 64% 0.00 l 1.80 j 16.40 ij 26.10 ij
2 Carbendazim 12% + Mancozeb 63% WP 100a 100 a 100 a 100 a
3 Carbendazim 50% WP 100a 100 a 100 a 100 a
4 Mancozeb 50% + Carbendazim 25% WS 94.40 b 100a 100 a 100 a
5 Mancozeb 75% WP 23.30 j 28.05 i 39.10 h 52.70 fg
6 Metalaxyl 35% WS 17.20 k 41.10 h 76.30 d 88.80 c
7 Chlorothalonil 75% WP 48.80g 54.40f 62.50e 72.50d
8 Propiconazole 25% EC 100a 100 a 100 a 100 a
9 Zineb 68% + Hexaconazole 4% WP 76.4 d 88.8 c 100a 100 a
17

23. Sr.
No.
Systemic fungicides
Mycelial inhibition (%) Compatible/
Non -
compatible
250 ppm 500 ppm 750 ppm 1000 ppm
1 Azoxystrobin 23% W/W 0.00 0.00 0.00 0.00 C
2 Carbendazim 50% WP 100.00 100.00 100.00 100.00 N
3 Tricyclozole 75% WP 0.00 0.00 0.00 0.00 C
4 Hexaconazole 5% EC 60.95 100.00 100.00 100.00 N
5 Thiophenate Methyl 70% WP 63.46 100.00 100.00 100.00 N
6 Metalaxyl 35% WS 0.00 0.00 0.00 10.58 C
7 Propiconazole 25% EC 100.00 100.00 100.00 100.00 N
CD 1.601 4.362 4.362 2.33
SE(m) 0.539 1.468 1.468 0.784
CV 3.286 6.975 6.887 3.45
Table 2a: Effect of systemic fungicides on the mycelial growth of T. harzianum.
Karnataka, India Sonavane and Venkataravanappa (2017)
18

24. Sr.No.
Chemical name
Concentration (ppm)
Mycelial inhibition (%)
Compatible/
Non
compatible
1000 1500 2000
1 Mancozeb 75WP 0.00 (0.00) 5.88 (11.36) 7.76 (16.02) C
2 Chlorothalonil 75WP 86.27 (68.32) 87.15 (69.00) 85.56 (67.86) N
3 Dinocap 48 EC 21.56 (27.57) 36.02 (36.86) 36.19 (36.97) C
4 Sulphur 80WDG 0.00 (0.00) 0.00 (0.00) 7.76 (16.02) C
5 Copper oxychloride 50WP 0.00 (0.00) 13.72 (21.70) 21.56 (27.57) C
CD 2.686 8.946 15.099
SE(d) 1.256 4.184 7.062
SE(m) 0.888 2.958 4.993
CV 12.838 35.916 44.088
Table 2b: Effect of contact fungicides on the mycelial growth of T. harzianum.
Karnataka, India 19 Sonavane and Venkataravanappa (2017)

25. Sr.
No.
Fungicides
Colony diameter (mm) at
0.05 % 0.1 % 0.2 %
1 Copper oxychloride 50% WDP 54.44 48.00 40.22
2 Mancozeb 64% + Metalaxyl 8% WP 72.22 67.77 50.9
3 Carbendazim 12% + Mancozeb 63%
WP
0 0 0
4 Fenamidone 10% + Mancozeb 50%
WG
60.00 50.9 34.44
5 Mancozeb 75% WP 72.22 40.00 15.00
6 Control 90 90 90
Table 3: Compatibility of Trichoderma viridae with fungicides
Nandini et al., (2018)
Andhra Pradesh
20

26. Andhra Pradesh Nandini et al., (2018)
Fig. 1: Mycelial growth of Trichoderma viridae at different concentration with different fungicides
21

27. Sr.
No.
Fungicides
Colony diameter (mm) of T1 at C.D.
(5%)
50 ppm 100 ppm 250 ppm 500 ppm
1 Mancozeb 75% WP 85.00 63.75 47.75 31.75 2.62
2 Thiram 75% WP 81.50 79.50 63 32.50 1.79
3 Chlorothalonil 75% WP 73.00 62.50 48.25 24.75 1.55
4 Carbendazim 50% WP 0.00 0.00 0.00 0.00 -
5 Propiconzole 25% EC 0.00 0.00 0.00 0.00 -
6 Tridemorph 75% EC 23.00 14.25 10.25 4.50 0.90
7 Thiophanate methyl 70% WP 25.75 16.75 6.00 0.00 1.08
Table 4: Compatibility of Trichoderma viride isolate T1 with fungicides
Jabalpur, M.P Kumar et al., (2019)
22

28. Treatment Concentration(ppm) Radial growth of Th-8 isolate
(mm)
Inhibition over control
(%)
Thiram
250 39.33 56.29 (48.59)
500 25.67 71.48 (57.49)
1000 17.67 80.37 (63.67)
Copper oxychloride
250 89.00 1.11 (6.09)
500 87.50 2.77 (11.85)
1000 82.33 8.52 (16.94)
Mancozeb
250 88.33 1.86 (6.35)
500 85.50 5.00 (12.89)
1000 78.33 12.97 (21.24)
Metalaxyl
250 89.33 0.75 (4.02)
500 87.66 2.60 (8.63)
1000 84.0 6.67 (14.91)
Control 90.0 0.00 (0.00)
C.D. (P<0.5) 2.04 (1.62)
SE(m)± 0.71 (0.42)
Table 5: Evaluation of some commonly used fungicides for their compatibility with Trichoderma
harzianum in vitro
Maurya et al., (2020)
Bihar, India 23

29. Bihar, India
Fig. 2: Mycelial growth of Trichoderma harzianum at different concentration with different fungicides
24 Maurya et al., (2020)

30. Sr. No. Fungicides
Inhibition per cent #
Mean
Concentrations (ppm)
500 1000 1500 2000
1. Captan 50% WP 51.10 (45.65)* 75.92 (60.64) 86.66 (68.61) 93.70 (75.50) 76.85
2. Copper hydroxide 53.8% W/W 0.74 (4.94) 5.92 (14.1) 7.96 (16.40) 12.59 (20.79) 6.80
3. Copper oxychloride 50% WP 0.00 (0.00) 4.81 (12.68) 11.48 (19.82) 18.89 (25.77) 8.80
4. Mancozeb 80% WP 0.00 (0.00) 4.81 (12.68) 10.74 (19.14) 12.59 (20.79) 7.04
5. Propineb 70% WP 11.48 (19.81) 21.11 (27.37) 33.33 (35.28) 50.74 (45.45) 29.17
6. Control 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00
S.Em± CD at 1%
Fungicides (F) 0.32 1.19
Concentrations (C) 0.26 0.98
F X C 0.65 2.39
Table 6a: In vitro evaluation of contact fungicides on the growth of Trichoderma asperellum
Karnataka, India Maheshwary et al., (2020)
25

31. Sr. No. Fungicides
Inhibition per cent #
Mean
Concentrations (ppm)
5 25 50 100
1. Azoxystrobin 25% SC 10.37 (18.79) 17.77 (24.94) 22.96 (28.64) 38.14 (38.16) 22.31
2. Metalaxyl 35% WS 0.00 (0.00)* 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00
3. Tebuconazole 25.9% EC 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
4. Propiconazole 25% EC 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
5. Carbendazim 50% WP 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
6. Control 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00
S.Em± CD at 1%
Fungicides (F) 0.08 0.31
Concentrations (C) 0.07 0.25
F X C 0.16 0.61
Table 6b: In vitro evaluation of systemic fungicides on the growth of Trichoderma asperellum
Maheshwary et al., (2020)
Karnataka, India 26

32. Sr. No. Fungicides
Inhibition per cent #
Mean
Concentrations (ppm)
1000 1500 2000 2500
1 Tebuconazole 50% + Trifloxystrobin 25% WG 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
2 Azoxystrobin 18.2% +Difenoconazole 11.4% SC 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
3 Metalaxyl-M 4% + Mancozeb 64% WP 0.00 (0.00)* 0.00 (0.00) 0.00 (0.00) 12.03 (20.31) 3.01
4 Carbendazim12% + Mancozeb 63% WP 100 (90.05) 100 (90.05) 100 (90.05) 100 (90.05) 100
5 Control 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00
S.Em± CD at 1%
Fungicides (F) 0.09 0.34
Concentrations (C) 0.08 0.31
F X C 0.18 0.69
Table 6c: In vitro evaluation of combination fungicides on the growth of Trichoderma asperellum
Maheshwary et al., (2020)
Karnataka, India 27

33. Sr.
No.
Fungicides Concentration (%)
Growth of Mycelium
(mm)
Per cent decrease over
control
Per cent
compatibility
1. Carbendazim
0.10 35.00 d 61.11 38.89
0.20 23.00 f 74.44 25.56
0.30 14.00 g 84.44 15.56
2. Copper oxychloride
0.20 49.00 b 45.55 54.45
0.30 28.00 e 68.88 31.12
0.40 17.00 g 81.11 18.89
3. Thiophanate methyl
0.05 16.00 g 82.22 17.78
0.10 15.00 g 83.33 16.67
0.20 14.00 g 84.44 15.56
4. Benomyl
0.05 10.00 h 88.88 11.12
0.10 9.00 h 90.00 10.00
0.20 8.00 h 91.11 8.89
5. Sodium hypochlorite
5.00 40.00 c 55.55 44.45
10.00 38.00 cd 57.77 42.23
15.00 23.00 f 74.44 25.56
6. Control - 90.00 a - -
Table 7: Effect of different fungicides on the radial mycelial growth of Trichoderma viride
Tamil Nadu, India Karpagavalli et al., (2020)
28

34. Fungicide
Per cent mycelial inhibition (concentration in ppm formulation basis)
50 100 250 500 1000 1500
Carbendazim
(50% WP)
100.00
(10.05) *
100.00
(10.05)
100.00
(10.05)
100.00
(10.05)
100.00
(10.05)
100.00
(10.05)
Carboxin
(75% WP)
6.89
(2.81)
22.89
(4.89)
41.11
(6.49)
55.33
(7.51)
81.56
(9.09)
100.00
(10.05)
Copper oxychloride
(50% WP)
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
51.33
(7.23)
55.33
(7.51)
Tebuconazole
(2% DS)
69.56
(8.40)
87.78
(9.42)
90.67
(9.57)
100.00
(10.05)
100.00
(10.05)
100.00
(10.05)
Control
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
0.00
(1.00)
CD(p=0.05) 0.109 0.081 0.047 0.037 0.056 0.037
* The figures in parentheses are square root transformed values
Table 8: Mycelial growth inhibition (%) of Trichoderma sp.-2 by different fungicides
Patel & Singh, (2020)
Palampur, HP 29

35. Agrochemicals and Concentrations (ppm)
Per cent growth inhibition
1 2 3 4 Mean
Copper oxychloride @1000, 1500, 2000 and 2500
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
Carbendazim @50, 100, 250 and 500
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
Mancozeb 50% + Carbendazim 25% @50, 100,
250 and 500
0.00
(0.00)
0.00
(0.00)
76.37
(94.44)
76.37
(94.44)
38.18
(38.21)
Methyl-o-demeton @ 250, 500, 1000 and 1500
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
Chlorpyrifos @ 250, 500, 1000 and 1500
50.81
(60.06)
55.25
(67.50)
58.91
(73.34)
65.35
(82.61)
57.58
(71.26)
Cartap hydrochloride @ 250, 500, 1000 and 1500
57.88
(71.73)
61.72
(77.55)
64.44
(81.39)
71.50
(89.93)
63.88
(80.62)
Quizalofop-ethyl @ 500, 1000, 1500 and 2000
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
76.37
(94.44)
Pendimethalin @ 500, 1000, 1500 and 2000
55.76
(68.34)
62.98
(79.37)
63.05
(79.46)
75.92
(94.08)
64.43
(81.37)
Mean
51.88
(61.90)
55.48
(67.88)
63.73
(80.41
66.63
(84.27)
Table 9 : Compatibility of promising agrochemicals with promising fungal bioagent Trichoderma
harzianum (in vitro)
JAU, Junagadh Vyas et al., (2020)
30
Values outside parenthesis are arcsine transformed and inside the parentheses, are re-transformed values

36. S. No. Fungicide/ Treatments Concentrations
Radial growth
(mm) 24 h
Radial growth
(mm) 48 h
Radial growth
(mm) 72 h
Radial growth
(mm) 96h
1.
Hexaconazole
10ppm 0.00 0.00 6.00 10.00
2. 15ppm 0.00 0.00 0.00 8.00
3. 20ppm 0.00 0.00 0.00 6.00
4.
Tebuconazole &
Trifloxystrobin
10ppm 0.00 8.00 12.00 22.00
5. 15ppm 0.00 6.00 10.00 18.00
6. 20ppm 0.00 0.00 0.00 6.00
7.
Propiconazole
10ppm 0.00 4.00 10.00 16.00
15ppm 0.00 0.00 8.00 12.00
8.
9. 20ppm 0.00 0.00 4.00 10.00
10.
Tebuconazole
10ppm 0.00 4.00 6.00 16.00
11. 15ppm 0.00 0.00 0.00 8.00
12. 20ppm 0.00 0.00 0.00 4.00
13. Control - 30.00 52.00 68.00 86.00
SE(m) 0.252 0.444 0.597 0.804
C.D. 0.728 1.282 1.724 2.32
Table 10: Effect of different concentrations of fungicides on radial growth of Trichoderma
harzianum strain IRRI-1 at 24, 48, 72 and 96 h of their inoculation
UP, India Singh et al.,(2021)
31

37. Fungicides tolerance in Pseudomonas fluorescens
32

38. Sr.
No.
Fungicides
Dose
g or ml/Lit
OD (610 nm) & Growth
on solid media
1 Hexaconazole 5% EC 1 ml 0.46b
2 Bordeaux mixture 1 % 0.28d
3 Copper oxychloride 50% WP 2 g 0.11e
4 Copper hydroxide 77% WP 2 g 0.09e
5 Carbendazim 50% WP 1 g 0.39c
6 Fosetyl- Al 80% WP 1 g 0.07e
7 Captan 70% WP+ Hexaconazole 5% WP 2 g 0.28d
8 Control 0.54a
CD (0.05) = 0.06; CV% = 11
Table 11: In vitro compatibility of fungicides against P. fluorescens
Kerala, India Dhanya et al. (2016)
33

39. Sr.
No.
Fungicides
Concentration (%)
0.05 0.1 0.2 0.3
1 Propiconazole 25% EC + + + + + + + + + + + +
2 Hexaconazole 5% EC + + + + + + + + + + + +
3 Tebuconazole 25.9% EC + + + + + + + + + + + +
4 Azoxystrobin 23 % SC + + + + + + + + + + + +
5 Copper oxychloride 50% WP + - - -
6 Mancozeb 75% WP + + - -
7 Control + + + + + + + + + + + +
(-) : No growth; (+) : Poor growth; (++) : Moderate growth; (+++) : Good growth
Louis et al. (2016)
Kerala, India
Table 12: Compatibility of Pseudomonas fluorescens with selected fungicides
34

40. Sr.
No.
Fungicides
Diameter of inhibition zone (mm) at
0.05 % 0.1 % 0.2 %
1 Copper oxychloride 50% WDP 10.00 12.55 13.22
2 Mancozeb 4% + Metalaxyl 64% W/W 0 0 0
3 Carbendazim 12% + Mancozeb 64% WP 0 0 0
4 Fenamidone 10% + Mancozeb 50% WG 0 9 12.55
5 Mancozeb 75% WP 0 0 0
6 Control 0 0 0
Table 13: Compatibility of Pseudomonas flourescens with fungicides
Andhra Pradesh Nandini et al., (2018)
35

41. Mancozeb+ Metalaxyl Copper oxychloride Fenamidone 10% +
Mancozeb 50%
Andhra Pradesh Nandini et al., (2018)
36

42. Sr. No. Fungicides
Growth of P. fluorescens 8
100 ppm 500 ppm 1000 ppm
1 Propiconazole 25 EC + + + + + +
2 Tebuconazole 43 SC + + + + + +
3 Trifloxystrobin 25%+ tebuconazole 50% WG + + + + + +
4 Azoxystrobin 23% SC + + + + + + + + +
5 Carbendazim 50% WP + + + + + +
6 Carbendazim 12% + mancozeb 63% WP + + + + + + + +
7 Control + + + + + + + + +
Table 14 : Compatibility of Pseudomonas fluorescens isolate with commonly used fungicides
Hanuman and Bindu (2018)
Guntur 37
(+) : Poor growth; (++) : Moderate growth; (+++) : Good growth

43. Azoxystrobin
Carbendazim + mancozeb
Trifloxistrobin + tebuconazole
Guntur Hanuman and Bindu(2018)
38

44. Agrochemicals and their concentrations in ppm
Per cent growth inhibition
1 2 3 4 Mean
Copper oxychloride@1000, 1500, 2000 and 2500
17.5
(8.89)
17.5
(8.89)
17.5
(8.89)
17.5
(8.89)
17.5
(8.89)
Carbendazim@50, 100, 250 and 500
15.95
(7.55)
15.95
(7.55)
15.95
(7.55)
15.95
(7.55)
15.95
(7.55)
Mancozeb 50% +Carbendazim 25%@50, 100, 250 and 500
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
Methyl-o-demeton@ 250, 500, 1000 and 1500
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
Chlorpyrifos@ 250, 500, 1000 and 1500
14.30
(6.11)
14.30
(6.11)
14.30
(6.11)
14.30
(6.11)
14.30
(6.11)
Cartap hydrochloride @ 250, 500, 1000 and 1500
16.20
(7.78)
16.20
(7.78)
16.20
(7.78)
16.20
(7.78)
16.20
(7.78)
Pendimethalin @ 500, 1000, 1500 and 2000
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
13.64
(5.56)
Mean
13.45
(5.41)
13.45
(5.41)
13.45
(5.41)
13.45
(5.41)
13.45
(5.41)
Agrochemicals (A) Conc. (C) A x C
S. Em. ± 0.17 0.06 0.35
C. D. at 5% 0.49 0.17 NS
C.V.% 4.52
Table 15: Compatibility of promising agrochemicals with promising fungal bio agent Pseudomonas
fluorescens (in vitro)
JAU, Junagadh Vyas et al., (2020)
39

45. Sr.No.
Fungicide/
Treatmens Concentration
CFUS
after 24hrs
Percent
Inhibition
CFUS
after 48hrs
Percent
Inhibition
CFUS
after 72
hrs
Percent
Inhibition
CFUS
after 96hrs
Percent
Inhibition
1.
Hexaconazole 10ppm. 0.00 100 3.00 89.66 7.00 83.33 13.00 80.88
Nativo 10ppm. 0.00 100 8.00 72.41 18.00 57.14 22.00 67.65
Propiconazole 10ppm. 0.00 100 4.00 86.21 11.00 73.81 16.00 76.47
Tebuconazole 10ppm. 0.00 100 7.00 75.86 9.00 78.57 14.00 79.41
2.
Hexaconazole 15ppm. 0.00 100 0.00 100.00 5.00 88.10 12.00 82.35
Nativo 15ppm. 0.00 100 5.00 82.76 11.00 73.81 17.00 75.00
Propiconazole 15ppm. 0.00 100 0.00 100.00 6.00 85.71 13.00 80.88
Tebuconazole 15ppm. 0.00 100 0.00 100.00 0.00 100.00 9.00 86.76
3.
Hexaconazole 20ppm. 0.00 100 0.00 100.00 4.00 90.48 8.00 88.24
Nativo 20ppm. 0.00 100 0.00 100.00 3.00 92.86 6.00 91.18
Propiconazole 20ppm. 0.00 100 0.00 100.00 4.00 90.48 7.00 89.71
Tebuconazole 20ppm. 0.00 100 0.00 100.00 0.00 100.00 6.00 91.18
4.
Hexaconazole 25ppm. 0.00 100 0.00 100.00 2.00 95.24 4.00 94.12
Nativo 25ppm. 0.00 100 0.00 100.00 0.00 100.00 2.00 97.06
Propiconazole 25ppm. 0.00 100 0.00 100.00 0.00 100.00 5.00 92.65
Tebuconazole 25ppm. 0.00 100 0.00 100.00 0.00 100.00 3.00 95.59
Control 15.00 0 29.00 0.00 42.00 0.00 68.00 0.00
Table 16: In vitro compatibility of Pseudomonas fluorescens with different systemic fungicides
UP, India Singh et al.,(2021)
40

46. Sr.
no.
Treatments
Inhibition
Zone (mm) at
48 hrs.
Inhibition
Zone (mm) at
72 hrs.
Average
1 Carbendazim 50% WP @ 1000 ppm 0.00 0.00 0.00
2 Tebuconazole 25.9% EC @ 1000 ppm 8.75 10.12 9.43
3 Carboxin 37.5% + Thirum 37.5% WP @ 2000 ppm 16.25 18.12 17.18
4
Tebuconazole 50% + Trifloxystrobin 25% WG @ 2000
ppm
0.00 0.00 0.00
5 Carbendazim 25% + Mancozeb 50% WP @ 2000 ppm 0.00 0.00 0.00
6 Control 0.00 0.00 0.00
SE± 0.35 0.32
CD at 1% 1.01 0.94
Table 17. In vitro compatibility of fungicides with potential isolates of Pseudomonas fluorescens
Sahane et al.,(2021)
Maharashtra, India 41

47. Fungicides tolerance to Trichoderma spp. in vivo

48. Sr.
No.
Treatment
Disease Incidence (%)
Pooled (2012-13 to 2014-15)
Stem rot Collar rot Aflarot
1 ST of Trichoderma 31.29 (33.5) 8.33 (16.47) 1.84 (7.71)
2 ST of mancozeb75% WP 31.70 (33.94) 5.03 (12.79) 1.64 (7.27)
3 ST of carboxin 37.5 % + thirum 37.5 % 44.58 (41.67) 12.17 (20.31) 2.51 (8.90)
4 T2 +Seed treatment of Trichoderma 36.54 (36.77) 4.85 (12.44) 1.59 (6.96)
5 T3 +Seed treatment of Trichoderma 32.58 (34.27) 10.68 (18.29) 2.49 (8.88)
6
ST of carbendazim 12% WP+ mancozeb 63% WP + ST of
Trichoderma
30.31 (32.45) 2.42 (8.84) 1.58 (7.10)
7 ST of tebuconazole 2% DS + ST of Trichoderma 32.39 (33.90) 5.26 (13.02) 1.69 (7.39)
8 ST of thiram 75% SD + ST of Trichoderma 47.22 (42.93) 12.61 (20.69) 3.67 (10.9)
9 ST of imidacloprid 600 FS +ST of Trichoderma 43.62 (41.27) 9.05 (16.79) 2.42 (8.77)
10 Control 50.34 (45.28) 15.55 (22.27) 4.11 (11.5)
Table 18 : Compatibility of Trichoderma with different seed dressing agrochemicals used for the
management of diseases and pest in groundnut.
JAU, Junagadh Anon.(2015)
Figures in parenthesis are retransformed value 42

49. Sr. No. Treatments
Disease Incidence (%)
2012-13 2013-14 2014-15 Pooled
T1
Drenching of carbendazim 50%WP @ 0.05% one
month after sowing
11.18 (19.53) 8.83 (17.29) 12.93 (20.91) 11.13 (19.24)
T2
Three sprays of mancozeb 75% WP 0.25% at
35,50 and 65 DAS
7.11 (15.46) 12.75 (20.92) 8.87 (17.27) 9.62 (17.89)
T3
Three sprays of hexaconazole 5% EC 0.005% at
35,50 and 65 DAS
9.22 (17.67) 10.87 (19.25) 11.03 (19.22) 10.529(18.71)
T4
Soil application of T. harzianum one month
after germination of crop
4.09 (11.66) 5.66 (13.76) 5.77 (13.67) 5.28 (13.03)
T5
T1 + Soil application of T. harzianum one
month after germination of crop
3.60 (10.93) 5.22 (13.21) 5.33 (13.26) 4.81 (12.46)
T6
T2+ Soil application of T. harzianum one month
after germination of crop
5.16 (13.13) 5.91 (14.07) 6.87 (14.99) 6.11 (14.07)
T7
T3+ Soil application of T. harzianum one month
after germination of crop
4.27 (11.93) 6.76 (15.08) 6.07 (14.18) 5.77 (13.73)
T8
Control 12.71 (20.88) 14.27 (22.19) 14.53 (22.25) 13.94 (21.78)
Table 19 : Cumin wilt incidence under foliar application of fungicides and soil application of ‘Sawaj Trichoderma’
JAU, Junagadh Anon.(2015)
43

50. Table 20 : Evaluation of compatible fungicides with Trichoderma spp. in tomato crop under in vivo
conditions
Sr.
No.
Treatments
Concentration
(%)
Population density of
T. viride (Cfu g-1 soil)
Population density of
T. harzianum (Cfu g-1 soil)
30 DAT 60 DAT 30 DAT 60 DAT
1.
Mancozeb
(75% WP)
0.15 4.44 4.70 4.33 4.78
0.20 4.22 4.52 4.22 4.55
0.25 3.89 4.19 3.89 4.33
2.
Azoxystrobin
(23% SC)
0.05 4.78 4.93 4.33 5.00
0.10 4.67 4.89 4.22 4.89
0.15 4.00 4.33 3.89 4.78
3.
Metalaxyl
(35% WS)
0.15 3.89 4.30 3.67 4.45
0.20 3.67 4.00 3.56 4.22
0.25 3.22 3.63 3.33 3.89
4. Control - 5.33 5.44 5.22 5.44
S. Em± 0.24 0.20 0.28 0.27
CD @ 5% 0.71 0.58 0.82 0.80
44 Shashi kumar et al., (2019)
Karnataka,India

51. Some Research related to increase tolerance in
Trichoderma spp.

52.  Aqueous suspensions of conidia of Trichoderma harzianum wild strain WT-6 were
placed on juice agar and exposed to ultraviolet (UV) radiation for 100 min.
 Conidia of WT-6 from surviving colonies of the first irradiation were allowed to
germinate on the agar before a second exposure to UV for 100 min.
 The irradiated plates were incubated at 25 C under fluorescent light, and the resulting
colonies of T. harzianum were isolated and grown on the medium. Conidia from the
colonies that survived the second irradiation were placed on agar and UV-
irradiated for 100 min.
 Of 36 colonies that survived the three irradiations, 19 colonies from the third series
tolerated high concentration of benomyl (100-500 mg/ml) as indicated by growth
in solid and liquid media and conidial germination tests on benomyl-amended agar.
Evaluation of new biotypes of Trichoderma harzianum for tolerance to
benomyl and enhanced bio control capabilities.
45
USA Papavizas et al., (1982)

53. The UV-induced biotypes differed
considerably from WT-6 in
appearance, growth habit,
fungitoxic metabolite production
against a pathogen ability to survive
in soil.
Certain UV-induced biotypes that
were also tolerant to benomyl
suppressed the saprophytic activity
of Rhizoctonia solani in soil more
effectively than did the wild strain.
46

54. Induction of fungicide resistance for enhanced cellulase production in
Trichoderma harzianum
 In this study UV light was used to induced mutations in T. harzianum.
 Spores from one week old cultures were placed into coconut water
medium and then exposed to UV light for 50 minutes.
 Spores of surviving colonies were plated and exposed for 100 minutes.
Survivors were re-exposed for another 100 minutes.
 The best growing colonies that survived this treatment was screened
for resistance to benomyl, the active ingredient in the fungicide benlate.
 A highly resistant strain, HUV 250, was obtained. It survived up to 562
microgram benomyl/ml, a concentration three times that of the full
dosage recommended for this fungicide.
47
Sajise et al., (1994)
Phillippines

55.  The isolates that survived the first 50 and 100 minutes exposure to UV
and the parental strain do not grow even at 1 microgram/ml benomyl.
 Initial studies indicated that the benlate resistance was maintained even if
HUV 250 was kept at benlate-free medium.
 A comparison of the cellulases from crude enzyme extract obtained
through ammonium sulfate precipitation of the parental strain and those
of HUV 250 was done.
 Cellulase (CM-ase) and B-glucosidase activities of the new strain were
significantly higher than those of the parental.
 However, morphologically the two organisms are very similar.
48

56. Development of UV-induced Carbendazim-resistant mutants of
Trichoderma harzianum for integrated control of damping-off disease
 T. harzianum is very sensitive to carbendazim which is used as a seed-
dressing fungicide for the control of fungal pathogens.
 Hence in the present study, carbendazim-resistant mutants of T. harzianum
were developed through UV-irradiation.
 These mutants grew well in PDA medium containing the carbendazim even at
100 µg/ml.
 Sustained production of cellulase and chitinase by these mutants in the
presence of carbendazim in the growth medium was observed.
 Seed treatment with carbendazim followed by application of carbendazim-
resistant mutants of T. harzianum resulted in better plant stand, plant
biomass and less damping-off disease caused by R. solani in both
greenhouse and field conditions.
Jayraj and Radhakrishnan (2003)
TNAU, India 49

57. Metalaxyl (500 ppm)
Thiram(500 ppm)
Mancozeb (1000 ppm)
Copper oxycloride(1000 ppm)
Azoxystrobin(1000 ppm)
Sulphur 80 WDG(1500 ppm)
Conclusion
In Trichoderma spp.
Carbendazim (1000 ppm)
Mancozeb (2000 ppm)
Hexaconazole(3000 ppm)
Propiconazole(3000 ppm)
Tebuconazole(3000 ppm)
Azoxystrobin(3000 ppm)
In Pseudomonas spp.
Fungicides tolerance up to..
Incompatible with..
Carbendazim
Hexaconazole
Propiconazole
Tebuconazole
Copper oxycloride
Generally, Bacterial bio control agent Pseudomonas
spp. More tolerant to fungicides than Trichoderma spp.
Fungicides tolerance increased in
UV Induced mutants of Trichoderma spp.
50